bluuebunny/crossref_metadata_2025_split huggingface dataset:
P: 53 Extent of Sarcopenia Does Not Correlate With Degree of Minimal Hepatic Encephalopathy in patients on Treatment for Hepatric Encephalopathy
Authors: Rathi Sahaj , Montagnese Sara , Brocco Silvia , Zarantonello Lisa , Formentin Chiara , Mangini Chiara , Merkel Carlo , Angeli Paolo , Stramare Roberto , Amodio Piero 
Published: September 2019

BACKGROUND:
Minimal hepatic encephalopathy affects over half of the patients with cirrhosis. It leads to deterioration of quality of life due to cognitive deficit. Both presence of sarcopenia and portosystemic shunts have been implicated in development of minimal hepatic encephalopathy. We retrospectively assessed the significance of sarcopenia and presence of portosystemic shunts in patients with minimal hepatic encephalopathy.

METHODS:
Patients with cirrhosis of liver attending the liver clinic at University of Padua underwent a detailed elective outpatient assessment for neurocognitive function. This included Psychometric Hepatic Encephalopathy Score (PHES), computer based tests, and EEG. We then selected patients who underwent a computed tomography for any reason within 3 months of the neurocognitive assessment. The degree of sarcopenia and presence of portosystemic shunts was evaluated on the scan. Presence of more than one shunt was accounted and adjusted for appropriately using a physiological approach previously described.

RESULTS:
A total of 120 patients had a CT scan within 3 months of the neurocognitive assessment. Of these, 100 were found to be suitable for analysis. Mean age of the patients was 59 ± 9.6 yrs and 73% were males. Median Child Pugh score was 8 (IQR 6–10) and MELD was 12 (IQR 10–18). Alcohol and viral etiologies responsible for 40% and 39% cases respectively. Based on Psychometric Hepatic Encephalopathy Score (PHES), 39% had minimal hepatic encephalopathy. Medication details were available for 87 patients, 70% of whom were on some form of treatment of hepatic encephalopathy with lactulose, antibiotics, or probiotics. The degree of cognitive dysfunction (PHES) correlated significantly with education (r = 0.33), shunt size (r = −0.45), and degree of hyperammonemia (r = −0.48), but not with sarcopenia or degree of hepatic dysfunction. Correlation between total effective shunt size and PHES was more pronounced in women (r = −0.68). Upon multivariate analysis, none of these variables predicted PHES. EEG mean dominant frequency correlated significantly with total effective shunt size (r = −0.30), ammonia (r = −0.34), and Child-Pugh score (r = −0.38). None of the parameters significantly correlated with critical flicker frequency. There was no difference in the degree of sarcopenia among patients with and without MHE or history of overt HE. Moreover, degree of sarcopenia did not correlate with PHES, critical flicker frequency, ammonia level, or Child-Pugh score. Effective size of portosystemic shunt was higher in patients with MHE.

CONCLUSIONS:
Among patients with cirrhosis on anti-encephalopathy treatment, presence of sarcopenia does not correlate with neurocognitive parameters. Presence of large portosystemic shunts correlated with neurocognitive impairment.
P: 57 Thrombospondin-1 Worsens Azoxymethane-Induced Hepatic Encephalopathy Through Activation of Transforming Growth Factor Beta 1
Authors: Frampton Gabriel , Jefferson Brandi , Grant Stephanie , DeMorrow Sharon , McMillin Matthew 
Published: September 2019

BACKGROUND:
Acute liver failure (ALF) is a consequence of severe hepatic injury and is associated with poor clinical outcomes. Patients with ALF often present with neurological complications, called hepatic encephalopathy (HE). Transforming growth factor beta 1 (TGFβ1) is upregulated following liver damage and we have shown that TGFβ1 drives HE progression. Thrombospondin-1 (TSP-1) can activate latent TGFβ1 and therefore, we hypothesize that hepatic-derived TGFβ1 is activated by TSP-1, which exacerbates liver damage and HE associated with azoxymethane-induced ALF.

METHODS:
Male C57Bl/6 mice were treated with azoxymethane (AOM; 100 mg/kg BW) to induce ALF. Six hours after AOM injection, mice were injected with the TSP-1 antagonist LSKL (30 mg/kg) or SLLK as control. In parallel, male TSP-1 knockout mice and wild-type (WT) controls were injected with AOM. In AOM-treated mice, cognitive impairment was monitored by reflex and ataxia measurement. Liver histology was assessed by hematoxylin and eosin staining and serum transaminases were measured. Cleaved caspase 3 immunohistochemistry and TUNEL staining were used to assess apoptosis in liver tissue. Hepatic inflammation was determined by measuring IL-1β, IL-6 and TNFα expression via real-time PCR and ELISA assays. TGFβ1 and TSP-1 expression were assessed in liver, serum and cortex by immunoblotting, immunohistochemistry and real-time PCR. Cerebral edema and microglia activation were assessed and neuroinflammation was measured by assessing IL-1β, IL-6 and TNFα expression in the cortex.

RESULTS:
Mice injected with AOM had elevated hepatic, circulating and cortical TGFβ1 and TSP-1 levels, with the cortex only having elevated TGFβ1. LSKL-treated mice and TSP-1 knockout mice administered AOM had reduced activation of hepatic TGFβ1, hepatocyte apoptosis, inflammation, and hepatic injury compared to AOM and SLLK-treated mice or WT AOM-treated mice. LSKL-treated and TSP-1 knockout mice administered AOM had an increased latency to reach coma compared to SLLK-treated or WT mice. LSKL-treated mice and TSP-1 knockout mice had reduced TGFβ1 expression, less cerebral edema, attenuated microglia activation, and decreased expression of IL-1β, IL-6 and TNFα in the cortex compared to control mice.

CONCLUSIONS:
TGFβ1 and TSP-1 were elevated in the livers and serum of AOM-treated mice and strategies employed to reduce TSP-1 signaling reduced liver damage and neuroinflammation in the AOM mouse model of HE. Therefore, targeting TSP-1 signaling may be a novel therapeutic target for the management of both ALF and HE following acute liver injury.
P: 59 Supplementation With Synbiotics and/or Branched Chain Amino Acids in Hepatic Encephalopathy: A Pilot Randomised Placebo-Controlled Clinical Study
Authors: Vidot Helen , Cvejis Erin , Finegan Liam , Shores E. Arthur , Strasser Simone , McCaughan Geoffrey , Carey Sharon , Shackel Nicholas 
Published: September 2019

BACKGROUND:
Hepatic encephalopathy (HE) is common in patients with cirrhosis and characterised by reduced hepatic ammonia clearance. This is accompanied by alterations in gut bacteria and increased intestinal epithelial permeability that may be ameliorated with synbiotics (pro- and pre-biotics). Branched chain amino acids (BCAAs) are thought to have a role in the detoxification of ammonia. We investigated the effects of the administration of Synbiotics and/or BCAAs in treating HE.

METHODS:
Methods Participants with minimal HE (MHE) were randomised in an placebo-controlled study to receive synbiotics, BCAAs, or a combination of BCAAs and Synbiotics. The investigators were blinded to the supplements at all times. Relevant biochemical and nutritional data and depression and anxiety scores (DASS-21) were collected at entry, 4 weeks, and on completion, at 8 weeks. The Trail Making Test (TMT) and Inhibitory Control Test (ICT) were used to assess cognitive function in HE. Results were analysed using linear mixed effects regression analyses

RESULTS:
Sixty-one participants with MHE determined by the treating physician, confirmed on psychometric testing (TMT and ICT) and who were taking 63 ± 6 mLs lactulose/day were enrolled. Recruitment was limited by the widespread introduction of rifaximin in Australia during the recruitment period. The final intention to treat analysis included 49 participants who returned for at least 1 follow-up review. The mean age was 55.8 ± 6.1 years and 86% were males. Despite evidence of a placebo effect, there was significant improvement in TMT B (P ≤ 0.05) (Figure 1a) and ICT weighted lures (P = 0.007) (Figure 1b) in participants who received combined synbiotics/BCAAs treatment compared to placebo at study completion. Cognitive improvement occurred without a significant change in ammonia levels.

CONCLUSIONS:
To the best of our knowledge, this study is the first to report an improvement in cognitive function and, therefore, executive function in individuals with decompensated cirrhosis and MHE in response to oral supplementation with combined synbiotics and BCAAs. A larger study which includes measures of dysbiosis and intestinal epithelial permeability is needed to confirm these results.
P: 60 Characterization of a Novel Mouse Model of Type A Hepatic Encephalopathy
Authors: Williams Elaina , Grant Stephanie , Petrescu Anca , Frampton Gabriel , McMillin Matthew , DeMorrow Sharon 
Published: September 2019

BACKGROUND:
Type A hepatic encephalopathy (HE) describes the neurological complications caused by acute liver failure. Research into Type A HE has been hampered as existing models cause severe and rapid HE, with a limited therapeutic window and high mortality, that do not reflect all features observed in patients. The aim of this study was to establish and characterize an alternative mouse model of Type A HE that has a more protracted timeline of pathology.

METHODS:
Male C57Bl/6 mice were fed standard rodent chow enriched with 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine and 10% ammonium acetate (DDC + NH4). Neurobehavioral indices and neuromuscular deficits were assessed by open field test, rotarod, grip strength test and gait analysis. Serum, liver and brain tissue were collected after 13 days of DDC + NH4 feeding. Liver damage was assessed by serum chemistry and H&E staining. Serum and cortical ammonia and total bile acid content were assessed with colorimetric assays. Cerebral edema was assessed using the wet weight/dry weight method. Microglia activation was assessed by Iba1 immunofluorescence. The expression of proinflammatory cytokines were assessed by qPCR and EIA.

RESULTS:
DDC + NH4 feeding caused significant neurological and neuromuscular deficits in every test performed commencing after 5–7 days of feeding. The liver damage observed was like a cholestatic hepatitis phenotype of drug-induced liver injury in humans; AST, ALT, alkaline phosphatase and total bilirubin were elevated. Histologically, there was evidence of intrahepatic cholestasis, drug-induced bile duct blockage, ductular reaction and immune cell infiltration, with low incidence of hepatocyte necrosis and little to no discernible fibrosis. Serum and cortical ammonia and total bile acid levels were elevated, and there was increased brain water content, indicating cerebral edema in DDC + NH4 fed mice. Furthermore, there was evidence of microglia activation and increased proinflammatory cytokine expression in the cortex of DDC+NH4 group compared to control diet-fed mice.

CONCLUSIONS:
DDC + NH4 feeding generated key features of Type A HE, with a longer timeline of pathology than the current models. With further characterization, this model may prove an effective and preferable model for pre-clinical studies to identify the pathogenesis or possible treatment options for Type A HE.
P: 72 Hyponatremia Correction in Cirrhosis May Increase the Speed of Complex Information Processing
Authors: Watson Hugh , Guevara Monica , Gines Pere , Vilstrup Hendrik 
Published: September 2019

BACKGROUND:
Hyponatremia is frequent in patients with advanced cirrhosis and ascites. A decrease in plasma osmolality exacerbates astrocyte swelling and through this mechanism may worsen cognitive function. Our objective was to determine whether, in patients with cirrhosis and ascites, an increase in plasma sodium (spontaneous or induced by an aquaretic) leads to an improvement in cognitive function.

METHODS:
250 patients with liver cirrhosis and ascites were included in two double-blind, randomised, multicentre clinical studies of satavaptan (a vasopressin V2 antagonist). Patients were included after a "wash-out period" (one week on controlled sodium intake and spironolactone 100 mg/d) and received either placebo (n = 61) or satavaptan (n = 189) at three different doses (5, 12.5, 25 mg/d). Patients completed the trail-making test (TMT) at baseline and at 5–7 and 14–28 days of follow-up. The TMT (scored in seconds) assesses speed of information processing and is considered a test of simple (part A) and complex attention (part B). Improvement of hyponatraemia was defined as an increase in plasma sodium >5 mmol/l or to a value >135 mmol/l.

RESULTS:
At inclusion into the studies, patients with normonatremia exhibited better results on the TMT-A and TMT-B that could not be explained by differences on age. This group also exhibited better parameters of liver function (Table 1). Improvement of hyponatremia was associated with shortening on the time needed to complete TMT-B: −20 s vs −6.5 s (P = 0.02), but not TMT-A: −10 s vs −5 s (P = 0.16). Increase in serum osmolality was also associated with better TMT-B scores (P < 0.001 at Day 5-7 and P = 0.033 at Day 14–28), but this was not shown for TMT-A. Liver function parameters remained stable in both groups of patients. Improvement of hyponatremia at Day 5 was most commonly seen in patients that received satavaptan (59.7%, n = 82) than in those treated with placebo (18.5%, n = 28). Median changes in TMT scores with satavaptan (all doses pooled) compared to placebo were: TMT-A −10 s vs 0 s (P = 0.08) and TMT-B −20 s vs −7.5 s (P = 0.12).

CONCLUSIONS:
Improvement of hyponatremia in cirrhosis was associated with an increase in the speed of complex information processing. Improvement of hyponatremia was achieved more frequently with satavaptan treatment, which showed a trend towards better results on cognitive function.
P: 73 Acetate Attenuates the Astrocyte Swelling and Brain Edema in Severe Liver Failure
Authors: Moriyama Mitsuaki , Minamihata Tomoki , Jayakumar Arumugam R. , Norenberg Michael D. 
Published: September 2019

BACKGROUND:
Astrocyte swelling and brain edema are major complications of severe liver failure. Ammonia plays a major role in the development of astrocyte swelling/brain edema in this condition. However, current therapies have not thus far improved the outcome of liver failure induced astrocyte swelling/brain edema. Since acetate has been shown to have neuroprotective effect in other neurological conditions, likely though increased energy production, we examined whether acetate similarly protect cell swelling in cultured astrocytes post-ammonia exposure. We also examined whether treatment of rats with glyceryl triacetate (GTA), an acetate precursor, which is known to increase circulating, as well as tissue levels of acetate, alleviates the brain edema induced by the liver toxin thioacetamide (TAA).

METHODS:
Astrocyte cell volume was estimated by measuring the intracellular water space using 3-O-methylglucose. Brain water content was measured by using the gravimetric method.

RESULTS:
Exposure of astrocyte cultures to pathological concentrations of ammonia (NH4Cl; 5 mM) for 24 h significantly increased cell swelling. Co-treatment of ammonia with acetate reduced such swelling in a dose-dependent manner. Further, treatment of rats with TAA (250 mg/kg bw) for 3 days increased the brain water content, and that pretreatment of (intragastrically) TAA-treated rats with GTA (7.5 g/kg bw), attenuated brain edema.

CONCLUSIONS:
These findings strongly suggest that acetate supplementation will exert salutary effects in reducing brain edema in patients with severe liver failure.
P: 74 Cholestasis Decreases Dendritic Spine Density in a Rat Hippocampal Organotypic Culture
Authors: Giovannoni Laurianne , De Roo Mathias , Braissant Olivier , Henry Hugues , Cudalbu Cristina , Vutskits Laszlo , McLin Valérie 
Published: September 2019

BACKGROUND:
Executive functioning impairment in children with cholestatic liver disease is increasingly recognized. Injury to developing neuronal networks could be an underlying mechanism. Given that the CSF of rat pups following bile duct ligation is highly concentrated in ammonia, murocholic and taurocholic acids, we hypothesized that cholestasis may significantly alter density of synaptic contacts onto pyramidal neurons, known to be essential in the development of executive functioning.

METHODS:
Transverse hippocampal organotypic slice (400 µm thick) were harvested from 6–7-day-old Wistar rats. They were maintained for 15 days in a CO2-incubator (33°C). pc-DNA3.1-EGFP plasmid biolistic transfection was performed 7 days after harvesting. 3 days following transfection, control medium or experimental medium containing 100 µM α-murocholic-acid, 200 µM taurocholic-acid and 2.5 mM ammonium was added to the culture (day 0 of exposure, MIX condition). Confocal microscopy using imageJ was used for to manually count CA1 pyramidal neuron dendritic spines as proxies for excitatory synapses. Static analysis quantified dendritic spines density each day. Dynamic analysis quantified dendritic spines turnover (loss and neo-formation) for each 24 h time-window. Statistical analysis was conducted using PRISM software for multiple t-test or mixed-effect ANOVA.

RESULTS:
Static analysis showed a biphasic profile in MIX condition. During early phase (first 3 days of exposure), we observed >50% decrease in dendritic spine density compared to control (cf Figure 1, P < 0.001). On days 3 to 4), spine density recovered to reach control value. Dynamic analysis showed 15% loss in dendritic spines stability during the early phase of exposure to MIX condition, compared to controls, with comparable low rates of spine turnover. During the late phase of MIX exposure, spine turnover increased significantly in favor of spine neo-formation: spine neo-formation was 10 times higher (0.280 vs 0.023 spines.µm-1, P = 0.033) than controls, while spine loss was 6 times higher in neurons exposed to MIX (0.154 vs 0.026 spines.µm-1, P = 0.0026) than controls.

CONCLUSIONS:
We demonstrate here that mimicking cholestasis ex vivo leads to a biphasic response in spine density of rat hippocampal CA1 pyramidal neurons. Spine density decreases during the first 3-days of exposure to a mix of bile acids and ammonium chloride-owing to decreased in spine stability. This is followed by increased spine turnover with spine neo-formation in excess of spine loss, yielding a final spine density similar to control animals. Whether these new spines are functionally equivalent to controls remains to be determined.
P: 76 Reversible and Irreversible Neurological Complications in Hepatic Encephalopathy
Authors: Tong Xiaoying , Shamaladevi Nagarajarao , Ahmad Anis , Arcuri Jennifer , Norenberg Michael , Jayakumar Arumugam 
Published: September 2019

BACKGROUND:
Hepatic encephalopathy (HE) is a major neurological condition that occurs due to acute and chronic liver failure following drug toxicity, viral hepatitis, or exposure to various hepatotoxins. Acute HE (Type A HE) is associated with cerebral edema, increased intracranial pressure, coma and death. Chronic HE (Type C HE) is characterized by mental confusion, behavioral changes, and motor disturbances. It is currently unclear whether HE is reversible or irreversible. We therefore examined whether acute or chronic liver failure is reversible or irreversible in the thioacetamide (TAA) rat model of acute and chronic HE. Accordingly, rats were treated with the liver toxin TAA and brain edema, neurobehavioral, cognitive and motor deficits were measured.

METHODS:
Rats were treated with the liver toxin TAA and brain edema, neurobehavioral, cognitive and motor deficits were measured.

RESULTS:
Treatment of rats with high doses of TAA (to induce acute HE) led to brain edema within 2-3 days. Rats following TAA treatment were examined for longer time-periods. Rats did not exhibit any brain edema, although they expressed neurobehavioral, cognitive and motor deficits without any changes documented in blood and brain ammonia levels, as well as liver failure markers, as compared to normal rats. However, rats that underwent chronic liver failure for 10 days displayed neurobehavioral, cognitive and motor deficits, along with brain structural and molecular events, including reduced levels of astrocytic matricellular proteins, as well as reduced levels of neuronal proteins. We also found a reduction in synaptic density and in dendritic complexity. These changes correlated-well with increased blood/brain ammonia levels and with liver failure markers. However, when TAA treatment was withdrawn (after 10 days), and the rats monitored for longer time periods still showed neurobehavioral, cognitive and motor deficits, as well as defective neuronal integrity, even though blood and brain ammonia levels, as well as liver biochemical and molecular parameters were reversed.

CONCLUSIONS:
These findings strongly suggest that neurobehavioral, cognitive and motor deficits in HE cannot be reversed even when acute or chronic liver failure had been corrected.
P: 79 Central Inhibition of IGFBP3 Attenuates Symptoms of Hepatic Encephalopathy in a Mouse Model of Acute Liver Failure
Authors: Grant Stephanie , Williams Elaina , Petrescu Anca , Reinhart Evan , Blaney Hanna , Frampton Gabriel , McMillin Matthew , DeMorrow Sharon 
Published: September 2019

BACKGROUND:
Insulin-like growth factor binding protein 3 (IGFBP3) has a strong binding affinity for IGF proteins and can inhibit the function of IGF by preventing interaction with its receptor. In addition, IGFBP3 can exert IGF-independent effects via direct interaction with a putative IGFBP3 receptor. Little is known about the function of IGFBP3 in the brain, outside of a role in brain cancer development. We have recently demonstrated that IGF1 expression is suppressed in the frontal cortex in a number of rodent models of hepatic encephalopathy (HE) and central administration of recombinant IGF1 attenuates the neurological complications. However, the role of IGFBP3 in the pathogenesis of HE is unknown and is the aim of this study.

METHODS:
C57Bl/6 mice were injected with azoxymethane (AOM) to induce acute liver failure and HE. In parallel, mice were given an intracerebroventricular infusion of the IGFBP inhibitor, NBI31772 for 3 days prior to AOM injection. Cognitive impairment was monitored by reflex response assessment at various time points and neuromuscular deficits were assessed using a grip strength meter. Liver damage was determined by H&E staining and serum chemistry. The expression of IGFBP3 and IGF1 was assessed in the cortex by qPCR, EIA and immunofluorescence using NeuN (neuronal), GFAP (astrocytic) or Iba1 (microglial) antibodies as a counterstain. Microglia activation was evaluated by assessment of field Iba1 staining and microglia cell morphology. Neuroinflammation was also measured by the expression of key proinflammatory cytokines.

RESULTS:
IGFBP3 was predominantly expressed in neurons of the cortex and increased once significant neurological deficits were observed after AOM injection. Pretreatment of mice with the IGFBP inhibitor had no effect on the underlying liver damage, but delayed the onset of neurological symptoms, attenuated the neuromuscular deficits, inhibited neuroinflammation and increased the bioavailability of the neuroprotective IGF1 after AOM injection, suggesting that IGFBP3 may be contributing to the pathogenesis of HE.

CONCLUSIONS:
Elevated cortical IGFBP3 expression contributes to the pathogenesis of HE in AOM-treated mice via a mechanism that may involve increasing IGF1 bioavailability, although IGF1-independent effects can not be ruled out. Targeting IGFBP3 in the cortex may prove promising for the development of adjunct therapies to manage the neurological complications of acute liver failure.