Conclusion: These

Conclusion: These drug discovery findings indicate that autophagy plays a critical role in liver

regeneration and in the preservation of cellular quality, preventing hepatocytes from becoming fully senescent and hypertrophic. (Hepatology 2014;60:290–300) “
“Secretin stimulates ductal secretion by interacting with secretin receptor (SR) activating cyclic adenosine 3′,5′-monophosphate/cystic fibrosis transmembrane conductance regulator/chloride bicarbonate anion exchanger 2 (cAMPCFTRCl−/HCO AE2) signaling that is elevated by biliary hyperplasia. Cholangiocytes secrete several neuroendocrine factors regulating biliary functions by autocrine mechanisms. Melatonin inhibits biliary growth and secretin-stimulated choleresis in cholestatic bile-duct–ligated (BDL) rats by interaction with melatonin type 1 (MT1) receptor through down-regulation of cAMP-dependent signaling. No data exist regarding the role of melatonin synthesized locally by cholangiocytes in the autocrine regulation of biliary growth and function. In this study, we evaluated the (1) expression of arylalkylamine N-acetyltransferase (AANAT; the rate-limiting enzyme Adriamycin order for melatonin synthesis from serotonin) in cholangiocytes and (2) effect of local modulation of biliary AANAT expression on the autocrine proliferative/secretory responses of cholangiocytes. In the liver, cholangiocytes (and, to a lesser extent, BDL

hepatocytes) expressed AANAT. AANAT expression and melatonin secretion (1) increased in BDL, compared to normal rats and BDL rats treated with melatonin, and (2) decreased in normal and BDL rats treated with AANAT Vivo-Morpholino, compared to controls. The decrease in AANAT expression, and subsequent lower melatonin secretion by cholangiocytes, was associated with increased biliary proliferation and increased SR, CFTR, and Cl−/HCO AE2 expression. Overexpression of AANAT

in cholangiocyte cell lines decreased the basal proliferative rate and expression of SR, CFTR, and Cl−/HCO AE2 and ablated secretin-stimulated biliary secretion in these cells. Conclusion: Local modulation of melatonin synthesis may be important for management of the balance between biliary proliferation/damage find more that is typical of cholangiopathies. (HEPATOLOGY 2013) Cholangiocytes modify canalicular bile before it reaches the duodenum through a series of secretory/absorptive events regulated by gastrointestinal hormones, including secretin.1, 2 Secretin stimulates bile secretion by interaction with secretin receptor (SR; expressed only by large cholangiocytes in the liver).3 Binding of secretin to its receptor induces an increase in cyclic adenosine 3′,5′-monophosphate (cAMP) levels,1, 4 activation of protein kinase A (PKA), which results in the efflux of Cl− through the cystic fibrosis transmembrane conductance regulator (CFTR),4 and subsequent activation of the chloride bicarbonate anion exchanger 2 (Cl−/HCO AE2)5 stimulating bicarbonate secretion.

Conclusion: These

Conclusion: These Daporinad chemical structure findings indicate that autophagy plays a critical role in liver

regeneration and in the preservation of cellular quality, preventing hepatocytes from becoming fully senescent and hypertrophic. (Hepatology 2014;60:290–300) “
“Secretin stimulates ductal secretion by interacting with secretin receptor (SR) activating cyclic adenosine 3′,5′-monophosphate/cystic fibrosis transmembrane conductance regulator/chloride bicarbonate anion exchanger 2 (cAMPCFTRCl−/HCO AE2) signaling that is elevated by biliary hyperplasia. Cholangiocytes secrete several neuroendocrine factors regulating biliary functions by autocrine mechanisms. Melatonin inhibits biliary growth and secretin-stimulated choleresis in cholestatic bile-duct–ligated (BDL) rats by interaction with melatonin type 1 (MT1) receptor through down-regulation of cAMP-dependent signaling. No data exist regarding the role of melatonin synthesized locally by cholangiocytes in the autocrine regulation of biliary growth and function. In this study, we evaluated the (1) expression of arylalkylamine N-acetyltransferase (AANAT; the rate-limiting enzyme MAPK Inhibitor Library clinical trial for melatonin synthesis from serotonin) in cholangiocytes and (2) effect of local modulation of biliary AANAT expression on the autocrine proliferative/secretory responses of cholangiocytes. In the liver, cholangiocytes (and, to a lesser extent, BDL

hepatocytes) expressed AANAT. AANAT expression and melatonin secretion (1) increased in BDL, compared to normal rats and BDL rats treated with melatonin, and (2) decreased in normal and BDL rats treated with AANAT Vivo-Morpholino, compared to controls. The decrease in AANAT expression, and subsequent lower melatonin secretion by cholangiocytes, was associated with increased biliary proliferation and increased SR, CFTR, and Cl−/HCO AE2 expression. Overexpression of AANAT

in cholangiocyte cell lines decreased the basal proliferative rate and expression of SR, CFTR, and Cl−/HCO AE2 and ablated secretin-stimulated biliary secretion in these cells. Conclusion: Local modulation of melatonin synthesis may be important for management of the balance between biliary proliferation/damage selleck compound that is typical of cholangiopathies. (HEPATOLOGY 2013) Cholangiocytes modify canalicular bile before it reaches the duodenum through a series of secretory/absorptive events regulated by gastrointestinal hormones, including secretin.1, 2 Secretin stimulates bile secretion by interaction with secretin receptor (SR; expressed only by large cholangiocytes in the liver).3 Binding of secretin to its receptor induces an increase in cyclic adenosine 3′,5′-monophosphate (cAMP) levels,1, 4 activation of protein kinase A (PKA), which results in the efflux of Cl− through the cystic fibrosis transmembrane conductance regulator (CFTR),4 and subsequent activation of the chloride bicarbonate anion exchanger 2 (Cl−/HCO AE2)5 stimulating bicarbonate secretion.

This migration was observed in different rat strains, that is, AC

This migration was observed in different rat strains, that is, ACI6 and Lewis rats (Yu, unpublished data), indicating the essential nature of this subset. Because crude bone marrow cells also contain blood-borne migrating DCs,6 this subset can be isolated easily and might have potential for use in a DC vaccine. As expected, the proliferative response

in the parathymic-LN T-cell area after LT was considerably higher than in other secondary lymphoid organs. The maximal response in the LNs8 was as high as ∼2,500 BrdU+ cells/mm2 in the T-cell area (Fig. 4E), reflecting the additive effect of the CD172a+CD11b− and CD172a+CD11b+ migrating subsets through PD0332991 the direct allorecognition pathway. The diaphragmatic lymphatics provide major drainage for fluids or cells from the peritoneal cavity in many animals, including humans,21 by connecting to the regional LNs. In dogs, cats, rabbits, guinea pigs, and sheep, the mediastinal and/or parasternal LNs are draining LNs.14 In humans, the diaphragmatic lymphatics are connected to the anterior, right and left lateral, and posterior diaphragmatic

LN groups, which drain into the parasternal, posterior mediastinal, and brachiocephalic LNs.22 Therefore, after LT in both experimental and clinical settings, the LNs that drain the peritoneal cavity should be recognized as major sites of the intrahost T-cell response by immunogenic passenger DCs that migrate through the lymph, rather than the ordinary regional liver LNs. In the Irr(+) group, the intrahost T-cell response was restricted to the parathymic LNs, where the CD172a+CD11b+ subset formed clusters with recipient proliferating

cells HDAC activation from the beginning of the response. In the graft liver, the CD172a+CD11b+ subset made up the majority of DCs (Fig. 6C) and formed clusters similar to those involving the parathymic LNs (Supporting Fig. 1F). Therefore, we suggest that this subset induces a CD8+ T-cell response in vivo in the parathymic LNs and probably also in the graft through find more the direct allorecognition pathway in the Irr(+) group. Notably, in vitro experiments involving the mixed leukocyte reaction showed that the radioresistant CD172a+CD11b+ subset in the liver and hepatic lymph of donor rats induced an intense T-cell proliferative response comparable to the control splenic DCs (Fig. 7B). This subset constitutively expressed the B7-2 costimulatory molecules (CD86) and had up-regulated IL-2 receptor alpha (CD25) expression when isolated from the parathymic LNs (Fig. 2B) and the graft liver (Fig. 6D). Expression of a functional IL-2 receptor can be induced in mouse splenic and lung DCs as well as in Langerhans cells during maturation, and a synergistic effect of IL-2 on interferon-gamma production by DCs has been reported on.23 Taken together, these data demonstrate that this subset is functionally mature and possesses the strong allostimulating activity in vitro.

Methods: The clinical data of 93 patients with suspected small bo

Methods: The clinical data of 93 patients with suspected small bowel disease who underwent DBE from January 2008 to January 2013 in the 1st affiliated hospital of Guangxi Medical University were retrospectively

analyzed. Results: 98 DBE procedures were performed in 93 patients. 52 of them were performed by oral and 36 by anal route, 5 patients were underwent by both approaches. 51 case (54.8%) with small bowel lesions were detected by DBE examination, which were Nonspecific enteritis 14 case (27.5%), small bowl tumor 9 case (17.6%), crohn’s disease 7 case (13.7%), diverticulum 6 case (11.8%), ulcer 5 case (9.8%). The lesion detection rate of DBE, GSK126 purchase abdomen CT, CE, barium meal were 63.3%, 32.4%, 53.8%, 19.1%; The lesion detection rate of ≥60 years, <60 years old was 47.6%, 56.9%, which did not reach statistic difference. the lesion detection rate was 61.4% in obscure gastrointestinal bleeding, 37.5% in obscure abdominal pain and 69.2% in obscure diarrhea, which did not reach statistic difference. 1 case (1%) patient were perforation, No hemorrhage, pancreatitis, aspiration pneumonia or other serious complications happened. Conclusion: There is a high diagnosis and safety on DBE which is a useful tool to

diagnosis the small bowl diseases. Key Word(s): 1. double-balloon; 2. small bowl diseases; 3. clinical use; 4. safety; Caspase inhibitor review Presenting Author: YANG YU-LONG Additional Authors: TU QIU-YING Corresponding Author: YANG YU-LONG Affiliations: The Fourth Affiliated Hospital of Nanchang University;

The Second Hospital of Nanchang City Objective: With change of environment and lifestyle, incidence and prevalence of Crohn’s disease (CD) remains on upward trend in past decade. However, early detection of the disease is challenging, especially for the small bowel Crohn’s disease. This study aimed to investigate the effectiveness of capsule endoscopy in the early diagnosis of small bowel Crohn’s disease. Methods: A retrospective analysis of 67 patients with Crohn’s disease was conducted at the Fourth Affiliated Hospital of Nanchang University from March 2008 to March 2013. Clinic, radiographic, endoscopic and pathological findings were reviewed and compared to selleck screening library identify an early diagnostic approach of small bowel Crohn’s disease. Results: Of the patients studied, 31 were males and 36 were females, whose ages ranged from 18 to 78 years. The main clinical manifestations were abdominal pain, diarrhea, blood in stool, anemia, hypoalbuminemia, weight loss, fever, as well as oral ulcers, joint pain, and in some cases anal fissure, intestinal obstruction and/or other complications. The locations of disease were ileitis 54 (50.7%), colitis 12 (17.9%), and ileocolitis 11 (16.4%). Diagnostic efficacy of Barium contrast x-rays and colonoscopy was 25.0% (10/40) and 55.2% (37/67), respectively. In contrast, 70.7% (46/65) were positively diagnosed by capsule endoscopy, among which 38 had small bowel lesions while 8 were with colon lesions.

Methods: The clinical data of 93 patients with suspected small bo

Methods: The clinical data of 93 patients with suspected small bowel disease who underwent DBE from January 2008 to January 2013 in the 1st affiliated hospital of Guangxi Medical University were retrospectively

analyzed. Results: 98 DBE procedures were performed in 93 patients. 52 of them were performed by oral and 36 by anal route, 5 patients were underwent by both approaches. 51 case (54.8%) with small bowel lesions were detected by DBE examination, which were Nonspecific enteritis 14 case (27.5%), small bowl tumor 9 case (17.6%), crohn’s disease 7 case (13.7%), diverticulum 6 case (11.8%), ulcer 5 case (9.8%). The lesion detection rate of DBE, Small molecule library research buy abdomen CT, CE, barium meal were 63.3%, 32.4%, 53.8%, 19.1%; The lesion detection rate of ≥60 years, <60 years old was 47.6%, 56.9%, which did not reach statistic difference. the lesion detection rate was 61.4% in obscure gastrointestinal bleeding, 37.5% in obscure abdominal pain and 69.2% in obscure diarrhea, which did not reach statistic difference. 1 case (1%) patient were perforation, No hemorrhage, pancreatitis, aspiration pneumonia or other serious complications happened. Conclusion: There is a high diagnosis and safety on DBE which is a useful tool to

diagnosis the small bowl diseases. Key Word(s): 1. double-balloon; 2. small bowl diseases; 3. clinical use; 4. safety; selleck kinase inhibitor Presenting Author: YANG YU-LONG Additional Authors: TU QIU-YING Corresponding Author: YANG YU-LONG Affiliations: The Fourth Affiliated Hospital of Nanchang University;

The Second Hospital of Nanchang City Objective: With change of environment and lifestyle, incidence and prevalence of Crohn’s disease (CD) remains on upward trend in past decade. However, early detection of the disease is challenging, especially for the small bowel Crohn’s disease. This study aimed to investigate the effectiveness of capsule endoscopy in the early diagnosis of small bowel Crohn’s disease. Methods: A retrospective analysis of 67 patients with Crohn’s disease was conducted at the Fourth Affiliated Hospital of Nanchang University from March 2008 to March 2013. Clinic, radiographic, endoscopic and pathological findings were reviewed and compared to learn more identify an early diagnostic approach of small bowel Crohn’s disease. Results: Of the patients studied, 31 were males and 36 were females, whose ages ranged from 18 to 78 years. The main clinical manifestations were abdominal pain, diarrhea, blood in stool, anemia, hypoalbuminemia, weight loss, fever, as well as oral ulcers, joint pain, and in some cases anal fissure, intestinal obstruction and/or other complications. The locations of disease were ileitis 54 (50.7%), colitis 12 (17.9%), and ileocolitis 11 (16.4%). Diagnostic efficacy of Barium contrast x-rays and colonoscopy was 25.0% (10/40) and 55.2% (37/67), respectively. In contrast, 70.7% (46/65) were positively diagnosed by capsule endoscopy, among which 38 had small bowel lesions while 8 were with colon lesions.

32,33 This HCC cohort included 22 cases of small-sized (≤ 5 cm) H

32,33 This HCC cohort included 22 cases of small-sized (≤ 5 cm) HCC, 34 cases of medium-sized (5–10 cm) HCC, and 20 cases of large-sized (≥ 10 cm) HCC. According to the serum AFP level, the HCC cases could also fall into two groups: AFP ≤ 25 ng/mL (n = 31), and AFP > 25 ng/mL (n = 45). Venous blood, obtained after overnight fasting, was collected in the absence of anticoagulant into red-topped tubes, and allowed to clot for 30 min at 4°C. Serum was isolated from blood by centrifugation at 2500 g for 15 min at 4°C. Then the serum was aliquoted

and stored at −80°C until analysis. Blood samples from HCC patients were all drawn prior Selisistat purchase to initiation of HCC treatment. Serum clusterin concentrations were

measured using a commercially available sandwich ELISA according to the manufacturer’s instructions (Human Clusterin ELISA, Biovendor Laboratories Ltd, Modrice, Czech Republic). All assays were performed independently by the laboratory personnel who did not have clinical information, and each sample was assayed in duplicate. In the Biovendor Human Clusterin ELISA, standards, quality controls and diluted samples were incubated in microtitration wells pre-coated with monoclonal anti-human clusterin antibody. After 60 min and washing, biotin-labeled second monoclonal anti-human clusterin antibody was added and incubated with the captured see more clusterin for 60 min. After another washing, streptavidin-horse radish peroxidase

(HRP) conjugate was added. After 30 min incubation and the last washing step, the remaining conjugate was allowed to react with the substrate solution (TMB). The reaction was stopped by the addition of acidic solution (0.2 M H2SO4) and absorbance of the resulting yellow product was measured spectrophotometrically at 450 nm. The absorbance was proportional to the concentration of clusterin. A standard curve was constructed by plotting absorbance values versus clusterin concentrations of standards, and concentrations of unknown samples are determined using this standard curve. The serum clusterin and AFP values were reported as median (25–75th percentile). The descriptive statistics selleck chemicals were compared by box plots and then by anova. Differences between groups were evaluated by the Kruskal–Wallis test followed by Tamhane’s test. To determine the optimal cutoff value for clusterin and AFP in the diagnosis of HCC, receiver operating characteristic (ROC) curves were constructed using all possible cutoffs for each assay. The area under the curve (AUC) was calculated and compared. For sensitivity and specificity, 95% confidence intervals (CI) were also determined. A P-value of < 0.05 was considered statistically significant. All analyses were performed with SPSS 13.0 for windows (SPSS, Inc., Chicago, IL, USA). In this study, a total of 184 subjects were enrolled.

32,33 This HCC cohort included 22 cases of small-sized (≤ 5 cm) H

32,33 This HCC cohort included 22 cases of small-sized (≤ 5 cm) HCC, 34 cases of medium-sized (5–10 cm) HCC, and 20 cases of large-sized (≥ 10 cm) HCC. According to the serum AFP level, the HCC cases could also fall into two groups: AFP ≤ 25 ng/mL (n = 31), and AFP > 25 ng/mL (n = 45). Venous blood, obtained after overnight fasting, was collected in the absence of anticoagulant into red-topped tubes, and allowed to clot for 30 min at 4°C. Serum was isolated from blood by centrifugation at 2500 g for 15 min at 4°C. Then the serum was aliquoted

and stored at −80°C until analysis. Blood samples from HCC patients were all drawn prior Everolimus chemical structure to initiation of HCC treatment. Serum clusterin concentrations were

measured using a commercially available sandwich ELISA according to the manufacturer’s instructions (Human Clusterin ELISA, Biovendor Laboratories Ltd, Modrice, Czech Republic). All assays were performed independently by the laboratory personnel who did not have clinical information, and each sample was assayed in duplicate. In the Biovendor Human Clusterin ELISA, standards, quality controls and diluted samples were incubated in microtitration wells pre-coated with monoclonal anti-human clusterin antibody. After 60 min and washing, biotin-labeled second monoclonal anti-human clusterin antibody was added and incubated with the captured Selleck IBET762 clusterin for 60 min. After another washing, streptavidin-horse radish peroxidase

(HRP) conjugate was added. After 30 min incubation and the last washing step, the remaining conjugate was allowed to react with the substrate solution (TMB). The reaction was stopped by the addition of acidic solution (0.2 M H2SO4) and absorbance of the resulting yellow product was measured spectrophotometrically at 450 nm. The absorbance was proportional to the concentration of clusterin. A standard curve was constructed by plotting absorbance values versus clusterin concentrations of standards, and concentrations of unknown samples are determined using this standard curve. The serum clusterin and AFP values were reported as median (25–75th percentile). The descriptive statistics learn more were compared by box plots and then by anova. Differences between groups were evaluated by the Kruskal–Wallis test followed by Tamhane’s test. To determine the optimal cutoff value for clusterin and AFP in the diagnosis of HCC, receiver operating characteristic (ROC) curves were constructed using all possible cutoffs for each assay. The area under the curve (AUC) was calculated and compared. For sensitivity and specificity, 95% confidence intervals (CI) were also determined. A P-value of < 0.05 was considered statistically significant. All analyses were performed with SPSS 13.0 for windows (SPSS, Inc., Chicago, IL, USA). In this study, a total of 184 subjects were enrolled.

This study was supported by a grant from MEXT-Supported Program f

This study was supported by a grant from MEXT-Supported Program for the Strategic Research Foundation at Private Universities, 2012. “
“The severe shortage of donor liver for transplantation demands novel, improved methods of ex vivo preservation. Machine perfusion has the potential to not only recover livers that are currently unsuitable for transplantation, but also provide an opportunity to quantitatively assess the liver’s viability and serve as a platform

for pathology-specific intervention. In a recent proof-of-concept study we demonstrated that a subnor-mothermic machine perfusion (SNMP) system could support and improve the quality of human livers that were discarded for transplantation. In this work, 22 human livers were perfused with the purpose of characterizing the dynamics of livers during SNMP to elucidate the underlying metabolic mechanisms selleckchem by which machine perfusion recovers

marginal livers. Livers were perfused Panobinostat for 3 hours with Williams’ medium E at 21°C following standard procurement and clinically relevant cold ischemia (4-8 hours). Characteristics of the donor liver varied over selected parameters including warm ischemic time (WIT; 0 -54 min), macro- and microsteatosis (0-80%). Perfusion hydrodynamics, functional and injury markers were determined in the perfusion solution. The metabolic dynamics of SNMP were characterized by targeted metabolomic analysis of hourly time-course biopsies, see more identifying significant alterations for ∼150 primary metabolites and ∼300 lipid compounds, which were mapped onto a hepatic network model, revealed several canonical metabolic pathway modules. Briefly, SNMP appears to replete intracellular ATP content, with a 2.7-fold increase after 3 hours. Recovery is inferior in livers with increased macrosteatosis (>30%) as well as longer WIT (>30 min). Moreover, steatotic livers showed lower reduced glutathione (GSH:GSSG) at the end of perfusion, suggesting increased free radical formation. Overall, redox status improves during SNMP for all livers, reflected by NADPH:NADP and NADH:NAD ratios. The time-course dynamics of several

intracellular metabolites, such as uracil, show altered levels between high and low WIT groups pre-perfusion but intriguingly reach the same level post-perfusion, suggesting that SNMP metabolically conditions the organ to a more uniform steady state regardless of donor characteristics. Moreover, prolonged ischemia generally results in a reduction of TCA cycle intermediates pre-perfusion, which appear to increase again over the course of SNMP. These observations aid in understanding machine perfusion recovery mechanisms and pathology-specific identifiers and therapeutic targets. Ongoing work aims to develop multivariate metrics to provide comprehensive viability indicators and ex vivo recovery mechanisms. Disclosures: The following people have nothing to disclose: Bote G. Bruinsma, Gautham V. Sridharan, Pepijn D. Weeder, James H.

This study was supported by a grant from MEXT-Supported Program f

This study was supported by a grant from MEXT-Supported Program for the Strategic Research Foundation at Private Universities, 2012. “
“The severe shortage of donor liver for transplantation demands novel, improved methods of ex vivo preservation. Machine perfusion has the potential to not only recover livers that are currently unsuitable for transplantation, but also provide an opportunity to quantitatively assess the liver’s viability and serve as a platform

for pathology-specific intervention. In a recent proof-of-concept study we demonstrated that a subnor-mothermic machine perfusion (SNMP) system could support and improve the quality of human livers that were discarded for transplantation. In this work, 22 human livers were perfused with the purpose of characterizing the dynamics of livers during SNMP to elucidate the underlying metabolic mechanisms check details by which machine perfusion recovers

marginal livers. Livers were perfused Veliparib in vitro for 3 hours with Williams’ medium E at 21°C following standard procurement and clinically relevant cold ischemia (4-8 hours). Characteristics of the donor liver varied over selected parameters including warm ischemic time (WIT; 0 -54 min), macro- and microsteatosis (0-80%). Perfusion hydrodynamics, functional and injury markers were determined in the perfusion solution. The metabolic dynamics of SNMP were characterized by targeted metabolomic analysis of hourly time-course biopsies, click here identifying significant alterations for ∼150 primary metabolites and ∼300 lipid compounds, which were mapped onto a hepatic network model, revealed several canonical metabolic pathway modules. Briefly, SNMP appears to replete intracellular ATP content, with a 2.7-fold increase after 3 hours. Recovery is inferior in livers with increased macrosteatosis (>30%) as well as longer WIT (>30 min). Moreover, steatotic livers showed lower reduced glutathione (GSH:GSSG) at the end of perfusion, suggesting increased free radical formation. Overall, redox status improves during SNMP for all livers, reflected by NADPH:NADP and NADH:NAD ratios. The time-course dynamics of several

intracellular metabolites, such as uracil, show altered levels between high and low WIT groups pre-perfusion but intriguingly reach the same level post-perfusion, suggesting that SNMP metabolically conditions the organ to a more uniform steady state regardless of donor characteristics. Moreover, prolonged ischemia generally results in a reduction of TCA cycle intermediates pre-perfusion, which appear to increase again over the course of SNMP. These observations aid in understanding machine perfusion recovery mechanisms and pathology-specific identifiers and therapeutic targets. Ongoing work aims to develop multivariate metrics to provide comprehensive viability indicators and ex vivo recovery mechanisms. Disclosures: The following people have nothing to disclose: Bote G. Bruinsma, Gautham V. Sridharan, Pepijn D. Weeder, James H.

This study was supported by a grant from MEXT-Supported Program f

This study was supported by a grant from MEXT-Supported Program for the Strategic Research Foundation at Private Universities, 2012. “
“The severe shortage of donor liver for transplantation demands novel, improved methods of ex vivo preservation. Machine perfusion has the potential to not only recover livers that are currently unsuitable for transplantation, but also provide an opportunity to quantitatively assess the liver’s viability and serve as a platform

for pathology-specific intervention. In a recent proof-of-concept study we demonstrated that a subnor-mothermic machine perfusion (SNMP) system could support and improve the quality of human livers that were discarded for transplantation. In this work, 22 human livers were perfused with the purpose of characterizing the dynamics of livers during SNMP to elucidate the underlying metabolic mechanisms Fulvestrant supplier by which machine perfusion recovers

marginal livers. Livers were perfused LY2835219 cell line for 3 hours with Williams’ medium E at 21°C following standard procurement and clinically relevant cold ischemia (4-8 hours). Characteristics of the donor liver varied over selected parameters including warm ischemic time (WIT; 0 -54 min), macro- and microsteatosis (0-80%). Perfusion hydrodynamics, functional and injury markers were determined in the perfusion solution. The metabolic dynamics of SNMP were characterized by targeted metabolomic analysis of hourly time-course biopsies, selleckchem identifying significant alterations for ∼150 primary metabolites and ∼300 lipid compounds, which were mapped onto a hepatic network model, revealed several canonical metabolic pathway modules. Briefly, SNMP appears to replete intracellular ATP content, with a 2.7-fold increase after 3 hours. Recovery is inferior in livers with increased macrosteatosis (>30%) as well as longer WIT (>30 min). Moreover, steatotic livers showed lower reduced glutathione (GSH:GSSG) at the end of perfusion, suggesting increased free radical formation. Overall, redox status improves during SNMP for all livers, reflected by NADPH:NADP and NADH:NAD ratios. The time-course dynamics of several

intracellular metabolites, such as uracil, show altered levels between high and low WIT groups pre-perfusion but intriguingly reach the same level post-perfusion, suggesting that SNMP metabolically conditions the organ to a more uniform steady state regardless of donor characteristics. Moreover, prolonged ischemia generally results in a reduction of TCA cycle intermediates pre-perfusion, which appear to increase again over the course of SNMP. These observations aid in understanding machine perfusion recovery mechanisms and pathology-specific identifiers and therapeutic targets. Ongoing work aims to develop multivariate metrics to provide comprehensive viability indicators and ex vivo recovery mechanisms. Disclosures: The following people have nothing to disclose: Bote G. Bruinsma, Gautham V. Sridharan, Pepijn D. Weeder, James H.