A TDR was attached to the belt to position it as close to the dugong as possible. The length of the tether and the buoyancy of the cylinder allowed the satellite antenna to be exposed above the selleck compound water when a dugong was in water <3 m or swimming near the surface very slowly, thereby maximizing uplink opportunities. The whole tracking apparatus was retrieved as described in Sheppard et al. (2006). Data from TDRs were decoded using software provided by the manufacturer.

We used custom software to preprocess the data by identifying the level of the water surface (zero-offset) and removing dugong spikes, i.e., biologically implausible rapid changes in depth (Appendix S1A, Hagihara et al. 2011). Dive data collected within 5 min of a GPS and QFP fix were then subsampled (10 min in total, 5 min before and Selleckchem 3-Methyladenine after each fix, Appendix S1B). To avoid any potential postrelease behavioral responses, we only used data recorded ≥3 d after the day of tag deployment. However, no apparent changes in diving patterns were observed in the

3 d after deployment; capture and handling did not appear to trigger a flight response and dugongs stayed in the vicinity of the capture area (Sheppard et al. 2006; RH, unpublished data). Bathymetric models and tidal records (Maritime Safety Queensland, Department of Transport and Main Roads) were used to estimate the water depth at the time and geographic location for each fix. The bathymetric models were of 100 m resolution and generated by Sheppard (2008) in Hervey Bay and by Beaman (2010) in Moreton Bay. The depth at the location of each fix was identified by importing the bathymetric models and location fixes into ArcGIS 9.3.1 (Environmental MCE公司 Systems Research Institute 2009). Tidal heights were added or subtracted to the depth on the bathymetric charts to calculate the water depth experienced by the dugong at the time of each fix. Tidal range in Hervey Bay was ca. 4 m and in Moreton Bay 2.6 m during the deployment periods. We assumed

that estimated water depths remained constant for the 10 min around each fix. Previous experiments using dugong replicas found that the availability of dugongs varies with levels of turbidity and sea state (Pollock et al. 2006). Following Pollock et al. (2006), we examined the proportion of time dugongs spent in two detection zones: 0–1.5 m of the surface for turbid water and Beaufort sea state 3 (rougher conditions with very few whitecaps); and 0–2.5 m of the surface for clear water and sea state ≤2 (calm conditions with no whitecaps). We assigned “1” when a depth measurement was recorded within each of the detection zones and “0” when a depth measurement was recorded outside of the detection zone. The proportion of time dugongs spent in each detection zone was calculated by the sum of these numbers divided by the number of depth records.

A TDR was attached to the belt to position it as close to the dugong as possible. The length of the tether and the buoyancy of the cylinder allowed the satellite antenna to be exposed above the Pritelivir concentration water when a dugong was in water <3 m or swimming near the surface very slowly, thereby maximizing uplink opportunities. The whole tracking apparatus was retrieved as described in Sheppard et al. (2006). Data from TDRs were decoded using software provided by the manufacturer.

We used custom software to preprocess the data by identifying the level of the water surface (zero-offset) and removing dugong spikes, i.e., biologically implausible rapid changes in depth (Appendix S1A, Hagihara et al. 2011). Dive data collected within 5 min of a GPS and QFP fix were then subsampled (10 min in total, 5 min before and selleck products after each fix, Appendix S1B). To avoid any potential postrelease behavioral responses, we only used data recorded ≥3 d after the day of tag deployment. However, no apparent changes in diving patterns were observed in the

3 d after deployment; capture and handling did not appear to trigger a flight response and dugongs stayed in the vicinity of the capture area (Sheppard et al. 2006; RH, unpublished data). Bathymetric models and tidal records (Maritime Safety Queensland, Department of Transport and Main Roads) were used to estimate the water depth at the time and geographic location for each fix. The bathymetric models were of 100 m resolution and generated by Sheppard (2008) in Hervey Bay and by Beaman (2010) in Moreton Bay. The depth at the location of each fix was identified by importing the bathymetric models and location fixes into ArcGIS 9.3.1 (Environmental MCE公司 Systems Research Institute 2009). Tidal heights were added or subtracted to the depth on the bathymetric charts to calculate the water depth experienced by the dugong at the time of each fix. Tidal range in Hervey Bay was ca. 4 m and in Moreton Bay 2.6 m during the deployment periods. We assumed

that estimated water depths remained constant for the 10 min around each fix. Previous experiments using dugong replicas found that the availability of dugongs varies with levels of turbidity and sea state (Pollock et al. 2006). Following Pollock et al. (2006), we examined the proportion of time dugongs spent in two detection zones: 0–1.5 m of the surface for turbid water and Beaufort sea state 3 (rougher conditions with very few whitecaps); and 0–2.5 m of the surface for clear water and sea state ≤2 (calm conditions with no whitecaps). We assigned “1” when a depth measurement was recorded within each of the detection zones and “0” when a depth measurement was recorded outside of the detection zone. The proportion of time dugongs spent in each detection zone was calculated by the sum of these numbers divided by the number of depth records.

Results from the exposure–response analysis suggest that increasing filibuvir doses beyond those tested in studies 1 and 2 is unlikely to result in greater reductions in HCV RNA concentrations. Based on the relationship observed for filibuvir dose and exposure (data HM781-36B clinical trial not shown), doses in excess of 200 mg BID are expected to achieve 24-hour

exposures resulting in at least half the maximal response, whereas doses in excess of 600 mg BID are expected to achieve exposures approaching the maximal response. A phase 2a study evaluating the effect of filibuvir given at 200, 300, and 500 mg BID (given for 4 weeks in combination with pegIFN and RBV) on HCV RNA concentrations showed that a greater proportion of patients achieved rapid virological response (>60%) at all filibuvir doses tested compared with the standard of care (0%).21 The exposure–response analysis, in conjunction with phase 2a combination study results, indicates that doses producing at least half the maximal response in monotherapy studies for filibuvir may be sufficient when used in combination with pegIFN Dabrafenib and RBV to improve efficacy compared with current standard-of-care therapy. Variants at NS5B residue 423 provided a clear correlate

of virologic breakthrough in these clinical studies (P. Troke, M. Lewis, P. Simpson, K. Gore, J. Hammond, C. Craig, M. Westby, unpublished data, 2010).22 This finding is consistent with in vitro

medchemexpress resistance data,16, 22 where high-level resistance has been demonstrated with variants (isoleucine, threonine, and valine) at residue 423. This finding is also consistent with data reported for other Thumb 2 NNIs. Specifically, variants at position 423 were identified via clonal sequence analysis as being the most predominant following VCH-759 exposure.23 There is no scientific rationale to expect that cross-resistance would occur between filibuvir and protease inhibitors and polymerase inhibitors that bind in other pockets in the polymerase protein.24 Several studies have been conducted to investigate the prevalence of known HCV drug-resistance mutations, including Met423Thr/Val/Ile, in the untreated HCV-infected patient population.25, 26 According to these studies, variants at position 423 are present in 2%-3% of the untreated patient population, and are associated with a reduction in the replicative fitness of the virus. The impact of pretreatment position 423 variants and/or the reduced fitness associated with these variants on response to therapy was not evaluated and is thus not yet understood. Although no position 423 variants were detected at baseline in either of the filibuvir monotherapy studies, a novel variant (Arg422Lys) was detected at baseline in virus isolated from a patient in study 1. This patient achieved a <0.

Monday Basic Early Morning Workshops EMW-15 Macrophage Polarization Cell Cycle inhibitor in Liver Disease Laura E. Nagy, PhD and Costica Aloman, MD EMW-16 Animal Models

of PBC M. Eric Gershwin, MD and Juan F. Medina, MD, PhD EMW-17 Mechanisms of Drug-induced Liver Injury Hartmut Jaeschke, PhD and John J. Lemasters, MD, PhD EMW-18 Roles of Autophagy in Liver Pathophysiology Mark J. Czaja, MD and Wen-Xing Ding, PhD EMW-19 Inflammasome Involvement in Liver Disease Gyongyi Szabo, MD, PhD and Wajahat Z. Mehal, MD EMW-20 Role of MicroRNAs in Liver Disease Joshua Friedman, MD, PhD and Kalpana Ghoshal, PhD Monday Clinical Early Morning Workshops EMW-21 Controversies in Management of PSC Jayant A. Talwalkar, MD and Roger W. Chapman, MD, PhD EMW-22 Who and How to Screen for Wilson Disease Peter Ferenci, MD and Frederick K. Askari, MD, PhD EMW-23 Acute on Chronic Liver Failure: An Update K. Rajender Reddy, MD and Rajiv Jalan, Doxorubicin MD, PhD EMW-24 Emerging Therapies for Management of NASH Naga P. Chalasani, MD and Vlad Ratziu, MD EMW-25 Management of HIV/HCV Co-infection in the DAA Era Mark S. Sulkowski, MD and Richard K. Sterling, MD, MSc EMW-26 HCV: Management of Non-responders including DAA Marc G. Ghany, MD and Gregory T. Everson, MD EMW-27 Beneficial Effects of Coffee Consumption in Liver Disease Kiran Bambha, MD and Dawn M. Torres, MD EMW-28

Are Patients With Chronic Liver Disease At Greater Risk Of Drug-Induced Liver Injury? Paul B. Watkins, MD and Robert J. Fontana, MD Poster Session III Monday, MCE November 4 8:00 AM – 5:30 PM Hall E Refer to page 149 for Poster Presentations Exhibit Hall Monday, November 4 9:30 AM – 3:00 PM Hall D Plenary Session Presidential Plenary: Scientific Advances in Hepatology Monday, November 4 8:00 – 9:30 AM Hall E/General Session MODERATORS: Ronald J. Sokol, MD J. Gregory Fitz, MD 8:00 AM 103: Interleukin-33 Induces a Potent Cholangiocyte Proliferation via

a Novel Paracrine Circuit Jun Li, Pranavkumar Shivakumar, Stephanie Walters, Tatsuki Mizuochi, Reena Mourya, Kazuhiko Bessho, Jorge A. Bezerra 8:15 AM 104: Signaling via the osteopontin and high-mobility group box-1 axis drives the fibrogenic response to liver injury Elena Arriazu, Xiaodong Ge, Tung Ming Leung, Aritz Lopategi, Yongke Lu, Naoto Kitamura, Raquel Urtasun, Neil D. Theise, Natalia Nieto 8:30 AM 105: Cell Fate Tracking Reveals Hepatocytes as the Primary Cellular Source for Hepatocellular Carcinoma Regina Español Suñer, Xueru Mu, Christine Sempoux, Dianne H. Dapito, Frederic Lemaigre, Isabelle A. Leclercq, Robert Schwabe 8:45 AM 106: The mast cell stabilizer, cromolyn sodium, reduces bile duct ligated-induced biliary hyperplasia: a novel role for the in vivoparacrine influence of mast cells on biliary proliferation Laura Hargrove, Lindsey Kennedy, Taylor Francis, Kyle M. Hodges, Allyson B. Graf, Yoshiyuki Ueno, John F. Greene, Heather L.

Monday Basic Early Morning Workshops EMW-15 Macrophage Polarization Sorafenib purchase in Liver Disease Laura E. Nagy, PhD and Costica Aloman, MD EMW-16 Animal Models

of PBC M. Eric Gershwin, MD and Juan F. Medina, MD, PhD EMW-17 Mechanisms of Drug-induced Liver Injury Hartmut Jaeschke, PhD and John J. Lemasters, MD, PhD EMW-18 Roles of Autophagy in Liver Pathophysiology Mark J. Czaja, MD and Wen-Xing Ding, PhD EMW-19 Inflammasome Involvement in Liver Disease Gyongyi Szabo, MD, PhD and Wajahat Z. Mehal, MD EMW-20 Role of MicroRNAs in Liver Disease Joshua Friedman, MD, PhD and Kalpana Ghoshal, PhD Monday Clinical Early Morning Workshops EMW-21 Controversies in Management of PSC Jayant A. Talwalkar, MD and Roger W. Chapman, MD, PhD EMW-22 Who and How to Screen for Wilson Disease Peter Ferenci, MD and Frederick K. Askari, MD, PhD EMW-23 Acute on Chronic Liver Failure: An Update K. Rajender Reddy, MD and Rajiv Jalan, Venetoclax mw MD, PhD EMW-24 Emerging Therapies for Management of NASH Naga P. Chalasani, MD and Vlad Ratziu, MD EMW-25 Management of HIV/HCV Co-infection in the DAA Era Mark S. Sulkowski, MD and Richard K. Sterling, MD, MSc EMW-26 HCV: Management of Non-responders including DAA Marc G. Ghany, MD and Gregory T. Everson, MD EMW-27 Beneficial Effects of Coffee Consumption in Liver Disease Kiran Bambha, MD and Dawn M. Torres, MD EMW-28

Are Patients With Chronic Liver Disease At Greater Risk Of Drug-Induced Liver Injury? Paul B. Watkins, MD and Robert J. Fontana, MD Poster Session III Monday, 上海皓元医药股份有限公司 November 4 8:00 AM – 5:30 PM Hall E Refer to page 149 for Poster Presentations Exhibit Hall Monday, November 4 9:30 AM – 3:00 PM Hall D Plenary Session Presidential Plenary: Scientific Advances in Hepatology Monday, November 4 8:00 – 9:30 AM Hall E/General Session MODERATORS: Ronald J. Sokol, MD J. Gregory Fitz, MD 8:00 AM 103: Interleukin-33 Induces a Potent Cholangiocyte Proliferation via

a Novel Paracrine Circuit Jun Li, Pranavkumar Shivakumar, Stephanie Walters, Tatsuki Mizuochi, Reena Mourya, Kazuhiko Bessho, Jorge A. Bezerra 8:15 AM 104: Signaling via the osteopontin and high-mobility group box-1 axis drives the fibrogenic response to liver injury Elena Arriazu, Xiaodong Ge, Tung Ming Leung, Aritz Lopategi, Yongke Lu, Naoto Kitamura, Raquel Urtasun, Neil D. Theise, Natalia Nieto 8:30 AM 105: Cell Fate Tracking Reveals Hepatocytes as the Primary Cellular Source for Hepatocellular Carcinoma Regina Español Suñer, Xueru Mu, Christine Sempoux, Dianne H. Dapito, Frederic Lemaigre, Isabelle A. Leclercq, Robert Schwabe 8:45 AM 106: The mast cell stabilizer, cromolyn sodium, reduces bile duct ligated-induced biliary hyperplasia: a novel role for the in vivoparacrine influence of mast cells on biliary proliferation Laura Hargrove, Lindsey Kennedy, Taylor Francis, Kyle M. Hodges, Allyson B. Graf, Yoshiyuki Ueno, John F. Greene, Heather L.

The authors considered a number of interesting hypotheses.[10] Among them the most plausible includes the fact that some of the patients may have had a subclinical partial portal vein thrombosis at the study entry, which was undetected because Doppler ultrasonography of the abdomen was not performed at enrollment. Perhaps the sustained platelet increase following treatment with eltrombopag, risk factors such as an imbalance of coagulation,[16] portal hypertension Autophagy Compound Library screening and reduced blood flow, local inflammation or endothelial injury could have acted in combination to exacerbate subclinical portal vein thrombosis in these patients. Another interesting hypothesis

could be that platelets in cirrhosis are overactivated, as shown by the increased urinary excretion of markers of in vivo platelet activation observed in these patients. It is possible that the relatively rapid increase click here of the number of overactivated platelets may have acted as a trigger for thrombosis.[17] In conclusion, the study of Afdhal et al.[10] shows that the strategy of using eltrombopag in patients with cirrhosis undergoing elective invasive procedures is effective in increasing the platelet count and thus avoiding platelet transfusion, but carries

the risk of increasing the rate of thrombotic events. On the other hand, the benefit of increasing platelet counts in this population in order to prevent hemorrhagic events has not yet been established. Hence, the benefit of improving hemostasis at the expenses of increasing thrombotic risk should be carefully evaluated in individual patients. Armando Tripodi, Ph.D. “
“Background and Aim:  Studies on normal values of liver stiffness (LS) in subjects at “low risk” for liver disease are scant. The aim of the present study was to assess liver stiffness values in the subjects without overt liver disease with normal alanine aminotransferases (ALT)

and to determine potential factors, which may influence these 上海皓元 values with special reference to newly suggested updated upper limits of normal for ALT. Methods:  Liver stiffness measurements were performed in 445 subjects without overt liver disease (mean age, 41.1 ± 13.6; male, 73.5%) and normal liver enzymes. Results:  Mean LS value was 5.10 ± 1.19 kPa. LS values were higher in men than in women (5.18 ± 1.67 vs 4.86 ± 1.24 kPa, respectively, P = 0.008); in subjects with higher body mass index (BMI) category (Normal, overweight and obese subjects; 4.10 ± 0.75, 5.08 ± 0.66, and 6.05 ± 1.28 kPa, respectively; P < 0.001); in subjects with metabolic syndrome than in those without (5.63 ± 1.37 vs 5.01 ± 1.14 kPa, P = 0.001); and in subjects with ALT levels more than updated limits of normal compared to subjects with ALT levels less than updated limits of normal (5.68 ± 1.21 vs 4.77 ± 1.05 kPa, P < 0.001). On multiple linear regression, BMI and ALT was found to be significant predictor of LS.

The authors considered a number of interesting hypotheses.[10] Among them the most plausible includes the fact that some of the patients may have had a subclinical partial portal vein thrombosis at the study entry, which was undetected because Doppler ultrasonography of the abdomen was not performed at enrollment. Perhaps the sustained platelet increase following treatment with eltrombopag, risk factors such as an imbalance of coagulation,[16] portal hypertension Liproxstatin-1 cell line and reduced blood flow, local inflammation or endothelial injury could have acted in combination to exacerbate subclinical portal vein thrombosis in these patients. Another interesting hypothesis

could be that platelets in cirrhosis are overactivated, as shown by the increased urinary excretion of markers of in vivo platelet activation observed in these patients. It is possible that the relatively rapid increase Navitoclax supplier of the number of overactivated platelets may have acted as a trigger for thrombosis.[17] In conclusion, the study of Afdhal et al.[10] shows that the strategy of using eltrombopag in patients with cirrhosis undergoing elective invasive procedures is effective in increasing the platelet count and thus avoiding platelet transfusion, but carries

the risk of increasing the rate of thrombotic events. On the other hand, the benefit of increasing platelet counts in this population in order to prevent hemorrhagic events has not yet been established. Hence, the benefit of improving hemostasis at the expenses of increasing thrombotic risk should be carefully evaluated in individual patients. Armando Tripodi, Ph.D. “
“Background and Aim:  Studies on normal values of liver stiffness (LS) in subjects at “low risk” for liver disease are scant. The aim of the present study was to assess liver stiffness values in the subjects without overt liver disease with normal alanine aminotransferases (ALT)

and to determine potential factors, which may influence these MCE values with special reference to newly suggested updated upper limits of normal for ALT. Methods:  Liver stiffness measurements were performed in 445 subjects without overt liver disease (mean age, 41.1 ± 13.6; male, 73.5%) and normal liver enzymes. Results:  Mean LS value was 5.10 ± 1.19 kPa. LS values were higher in men than in women (5.18 ± 1.67 vs 4.86 ± 1.24 kPa, respectively, P = 0.008); in subjects with higher body mass index (BMI) category (Normal, overweight and obese subjects; 4.10 ± 0.75, 5.08 ± 0.66, and 6.05 ± 1.28 kPa, respectively; P < 0.001); in subjects with metabolic syndrome than in those without (5.63 ± 1.37 vs 5.01 ± 1.14 kPa, P = 0.001); and in subjects with ALT levels more than updated limits of normal compared to subjects with ALT levels less than updated limits of normal (5.68 ± 1.21 vs 4.77 ± 1.05 kPa, P < 0.001). On multiple linear regression, BMI and ALT was found to be significant predictor of LS.

The authors considered a number of interesting hypotheses.[10] Among them the most plausible includes the fact that some of the patients may have had a subclinical partial portal vein thrombosis at the study entry, which was undetected because Doppler ultrasonography of the abdomen was not performed at enrollment. Perhaps the sustained platelet increase following treatment with eltrombopag, risk factors such as an imbalance of coagulation,[16] portal hypertension selleck chemical and reduced blood flow, local inflammation or endothelial injury could have acted in combination to exacerbate subclinical portal vein thrombosis in these patients. Another interesting hypothesis

could be that platelets in cirrhosis are overactivated, as shown by the increased urinary excretion of markers of in vivo platelet activation observed in these patients. It is possible that the relatively rapid increase Obeticholic Acid molecular weight of the number of overactivated platelets may have acted as a trigger for thrombosis.[17] In conclusion, the study of Afdhal et al.[10] shows that the strategy of using eltrombopag in patients with cirrhosis undergoing elective invasive procedures is effective in increasing the platelet count and thus avoiding platelet transfusion, but carries

the risk of increasing the rate of thrombotic events. On the other hand, the benefit of increasing platelet counts in this population in order to prevent hemorrhagic events has not yet been established. Hence, the benefit of improving hemostasis at the expenses of increasing thrombotic risk should be carefully evaluated in individual patients. Armando Tripodi, Ph.D. “
“Background and Aim:  Studies on normal values of liver stiffness (LS) in subjects at “low risk” for liver disease are scant. The aim of the present study was to assess liver stiffness values in the subjects without overt liver disease with normal alanine aminotransferases (ALT)

and to determine potential factors, which may influence these MCE公司 values with special reference to newly suggested updated upper limits of normal for ALT. Methods:  Liver stiffness measurements were performed in 445 subjects without overt liver disease (mean age, 41.1 ± 13.6; male, 73.5%) and normal liver enzymes. Results:  Mean LS value was 5.10 ± 1.19 kPa. LS values were higher in men than in women (5.18 ± 1.67 vs 4.86 ± 1.24 kPa, respectively, P = 0.008); in subjects with higher body mass index (BMI) category (Normal, overweight and obese subjects; 4.10 ± 0.75, 5.08 ± 0.66, and 6.05 ± 1.28 kPa, respectively; P < 0.001); in subjects with metabolic syndrome than in those without (5.63 ± 1.37 vs 5.01 ± 1.14 kPa, P = 0.001); and in subjects with ALT levels more than updated limits of normal compared to subjects with ALT levels less than updated limits of normal (5.68 ± 1.21 vs 4.77 ± 1.05 kPa, P < 0.001). On multiple linear regression, BMI and ALT was found to be significant predictor of LS.

3A,E) that colocalized predominately, but not exclusively, with the iron storage protein, ferritin, in periportal regions of the liver (Supporting Fig. 2). The number of CD45+ inflammatory cells was significantly increased in the livers from Hfe−/−×Tfr2mut mice, compared with the other groups of mice (P < 0.05), whereas the number of CD45+ cells in Hfe−/−, Tfr2mut, and iron-loaded WT mice was not significantly different from those in non-iron-loaded WT mice (Fig. 3F). Another unique feature of Hfe−/− ×Tfr2mut mice was the evidence Pritelivir of inflammatory sideronecrosis of hepatocytes, which was not observed in any other group of mice (Fig. 3E). Liver injury

was assessed by examining plasma ALT as well as hepatic SOD and F2-isoprostane levels. Plasma ALT activity was increased in Hfe−/−×Tfr2mut mice by at least 1.8-fold, compared with all other types of mice (P < 0.001; Fig. 4A). Both hepatic copper/zinc (cytosolic) and manganese (mitochondrial) SOD activities were significantly decreased in all HH mice. In Hfe−/−×Tfr2mut mice copper/zinc SOD levels were similar, whereas manganese SOD levels were significantly lower than Hfe−/−

and Tfr2mut mice (P < 0.01; Fig. 4B). Liver F2-isoprostanes were elevated in all groups of HH mice, compared with non-iron-loaded WT mice (P < 0.01), with Hfe−/− ×Tfr2mut mice having similar liver F2-isoprostane levels to iron-loaded WT mice and significantly higher find more levels than either Hfe−/− or Tfr2mut mice (P < 0.01; Fig. 4C). Hepatic collagen deposition, a marker of fibrosis, was examined by histology using Sirius red and Masson's trichrome staining and by biochemical measurement of hydroxyproline levels. Hydroxyproline levels were increased

in all iron-loaded mice. In Hfe−/−×Tfr2mut mice, hydroxyproline levels were significantly increased, compared with Tfr2mut mice, and both were elevated, compared with Hfe−/− and iron-loaded WT mice (Fig. 4D; P < 0.05). Likewise, Hfe−/−×Tfr2mut mice had significantly increased Sirius red staining, compared with Hfe−/−, Tfr2mut, and iron-loaded WT mice (P < 上海皓元 0.05), which, in turn, exhibited greater collagen deposition than non-iron-loaded WT mice (P < 0.01; Fig. 5A-F). Sirius red staining revealed portal tract thickening and periportal fibrosis in Hfe−/−×Tfr2mut mice. In addition, there was evidence of portal tract bridging in Hfe−/− ×Tfr2mut mice, which was not evident in other groups. Quantification of Sirius red staining correlated with HIC (r2 = 0.98; P = 0.001), plasma NTBI (r2 = 0.82; P = 0.033), as well as hydroxyproline (r2 = 0.89; P = 0.015) and F2-isoprotane levels (r2 = 0.77; P = 0.048) in HH mice. This suggests that the collagen levels measured by biochemical assay were consistent with histological observations using Sirius red staining and were dependent on both plasma NTBI and HIC in HH mice. Furthermore, the intensity of trichrome staining, a commonly used, but less sensitive, marker of fibrosis, was also significantly enhanced in Hfe−/−×Tfr2mut and Tfr2mut mice (Fig.

3A,E) that colocalized predominately, but not exclusively, with the iron storage protein, ferritin, in periportal regions of the liver (Supporting Fig. 2). The number of CD45+ inflammatory cells was significantly increased in the livers from Hfe−/−×Tfr2mut mice, compared with the other groups of mice (P < 0.05), whereas the number of CD45+ cells in Hfe−/−, Tfr2mut, and iron-loaded WT mice was not significantly different from those in non-iron-loaded WT mice (Fig. 3F). Another unique feature of Hfe−/− ×Tfr2mut mice was the evidence Trametinib of inflammatory sideronecrosis of hepatocytes, which was not observed in any other group of mice (Fig. 3E). Liver injury

was assessed by examining plasma ALT as well as hepatic SOD and F2-isoprostane levels. Plasma ALT activity was increased in Hfe−/−×Tfr2mut mice by at least 1.8-fold, compared with all other types of mice (P < 0.001; Fig. 4A). Both hepatic copper/zinc (cytosolic) and manganese (mitochondrial) SOD activities were significantly decreased in all HH mice. In Hfe−/−×Tfr2mut mice copper/zinc SOD levels were similar, whereas manganese SOD levels were significantly lower than Hfe−/−

and Tfr2mut mice (P < 0.01; Fig. 4B). Liver F2-isoprostanes were elevated in all groups of HH mice, compared with non-iron-loaded WT mice (P < 0.01), with Hfe−/− ×Tfr2mut mice having similar liver F2-isoprostane levels to iron-loaded WT mice and significantly higher find more levels than either Hfe−/− or Tfr2mut mice (P < 0.01; Fig. 4C). Hepatic collagen deposition, a marker of fibrosis, was examined by histology using Sirius red and Masson's trichrome staining and by biochemical measurement of hydroxyproline levels. Hydroxyproline levels were increased

in all iron-loaded mice. In Hfe−/−×Tfr2mut mice, hydroxyproline levels were significantly increased, compared with Tfr2mut mice, and both were elevated, compared with Hfe−/− and iron-loaded WT mice (Fig. 4D; P < 0.05). Likewise, Hfe−/−×Tfr2mut mice had significantly increased Sirius red staining, compared with Hfe−/−, Tfr2mut, and iron-loaded WT mice (P < MCE公司 0.05), which, in turn, exhibited greater collagen deposition than non-iron-loaded WT mice (P < 0.01; Fig. 5A-F). Sirius red staining revealed portal tract thickening and periportal fibrosis in Hfe−/−×Tfr2mut mice. In addition, there was evidence of portal tract bridging in Hfe−/− ×Tfr2mut mice, which was not evident in other groups. Quantification of Sirius red staining correlated with HIC (r2 = 0.98; P = 0.001), plasma NTBI (r2 = 0.82; P = 0.033), as well as hydroxyproline (r2 = 0.89; P = 0.015) and F2-isoprotane levels (r2 = 0.77; P = 0.048) in HH mice. This suggests that the collagen levels measured by biochemical assay were consistent with histological observations using Sirius red staining and were dependent on both plasma NTBI and HIC in HH mice. Furthermore, the intensity of trichrome staining, a commonly used, but less sensitive, marker of fibrosis, was also significantly enhanced in Hfe−/−×Tfr2mut and Tfr2mut mice (Fig.