Search

The use of machine perfusion (MP) in liver transplantation (LT) is spreading worldwide. However, its efficacy has not been demonstrated, and its proper clinical use has far to go to be widely implemented. The Società Italiana Trapianti d’Organo (SITO) promoted the development of an evidence-based position paper. A 3-step approach has been adopted to develop this position paper. First, SITO appointed a chair and a cochair who then assembled a working group with specific experience of MP in LT. The Guideline Development Group framed the clinical questions into a patient, intervention, control, and outcome (PICO) format, extracted and analyzed the available literature, ranked the quality of the evidence, and prepared and graded the recommendations. Recommendations were then discussed by all the members of the SITO and were voted on via the Delphi method by an institutional review board. Finally, they were evaluated and scored by a panel of external reviewers. All available literature was analyzed, and its quality was ranked. A total of 18 recommendations regarding the use and the efficacy of ex situ hypothermic and normothermic machine perfusion and sequential normothermic regional perfusion and ex situ MP were prepared and graded according to the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) method. A critical and scientific approach is required for the safe implementation of this new technology.

Heart failure-either with reduced or preserved ejection fraction (HFrEF/HFpEF)-is a clinical syndrome of multi-factorial and gender-dependent aetiology, indicating the insufficiency of the heart to pump blood adequately to maintain blood flow to meet the body's needs. Typical symptoms commonly include shortness of breath, excessive fatigue with impaired exercise capacity, and peripheral oedema, thereby alluding to the fact that heart failure is a syndrome that affects multiple organ systems. Patients suffering from progressed heart failure have a very limited life expectancy, lower than that of numerous cancer types. In this position paper, we provide an overview regarding interactions between the heart and other organ systems, the clinical evidence, underlying mechanisms, potential available or yet-to-establish animal models to study such interactions and finally discuss potential new drug interventions to be developed in the future. Our working group suggests that more experimental research is required to understand the individual molecular mechanisms underlying heart failure and reinforces the urgency for tailored therapeutic interventions that target not only the heart but also other related affected organ systems to effectively treat heart failure as a clinical syndrome that affects and involves multiple organs.

During the early stages of the COVID-19 pandemic, EASL and ESCMID published a position paper to provide guidance for physicians involved in the care of patients with chronic liver disease. In the meantime, many countries and healthcare systems have been, or are still overwhelmed by the pandemic, significantly impacting on the care of this group of patients, whilst others have started to return towards their usual routine. In addition, many studies have been published focusing on how COVID-19 may affect the liver and how pre-existing liver diseases might influence the clinical course of COVID-19. While many aspects remain poorly understood, it has become increasingly evident that pre-existing liver diseases and liver injury during the course of the disease have to be kept in mind when caring for patients with COVID-19. Thus, this review should serve as an update on the previous position paper summarizing the evidence for liver disease involvement during COVID-19 and also provid, PNN was supported by the National Institute of Health Research (NIHR) Birmingham Biomedical Research Centre (BRC). The views expressed are those of the authors and not necessarily those of the NHS, the NIHR or the Department of Health.

The pervasive shortage of deceased donor liver allografts contributes to significant waitlist mortality despite efforts to increase organ donation. Ex vivo liver perfusion appears to enhance preservation of donor organs, extending viability and potentially evaluating function in organs previously considered too high risk for transplant. These devices pose novel challenges for organ allocation, safety, training, and finances. This white paper describes the American Society of Transplant Surgeons' belief that organ preservation technology is a vital advance, but its use should not change fundamental aspects of organ allocation. Additional data elements need to be collected, made available for organ assessment by transplant professionals to allow determination of organ suitability in the case of reallocation and incorporated into risk adjustment methodology. Finally, further work is needed to determine the optimal strategy for management and oversight of perfused organs prior to transplantation.

During the early stages of the coronavirus disease 2019 (COVID-19) pandemic, EASL and ESCMID published a position paper to provide guidance for physicians involved in the care of patients with chronic liver disease. While some healthcare systems are returning to a more normal routine, many countries and healthcare systems have been, or still are, overwhelmed by the pandemic, which is significantly impacting on the care of these patients. In addition, many studies have been published focusing on how COVID-19 may affect the liver and how pre-existing liver diseases might influence the clinical course of COVID-19. While many aspects remain poorly understood, it has become increasingly evident that pre-existing liver diseases and liver injury during the disease course must be kept in mind when caring for patients with COVID-19. This review should serve as an update on the previous position paper, summarising the evidence for liver disease involvement during COVID-19 and providing recommendations on how to return to routine care wherever possible. Yes

In this paper, we propose a generalized concept of microfluidic polydimethylsiloxane (PDMS) on paper (POP) devices, which combines well the merits of paper chips and PDMS chips. First, we optimized the conditions for accurate PDMS spatial patterning on paper, based on screen printing and a high temperature enabled superfast curing technique, which enables PDMS patterning to an accuracy of tens of microns in less than ten seconds. This, in turn, makes it available for seamless, reversible and reliable integration of the resulting paper layer with other PDMS channel structures. The integrated POP devices allow for both porous paper and smooth channels to be spatially defined on the devices, greatly extending the flexibility for designers to be able to construct powerful functional structures. To demonstrate the versatility of this design, a prototype POP device for the colorimetric analysis of liver function markers, serum protein, alkaline phosphatase (ALP) and aspartate aminotransferase (AST), was constructed. On this POP device, quantitative sample loading, mixing and multiplex analysis have all been realized.

New ultrasound methods that can be used to quantitatively assess liver fat content have recently been developed. These quantitative ultrasound (QUS) methods are based on the analysis of radiofrequency echoes detected by the transducer, allowing calculation of parameters for quantifying the fat in the liver. In this position paper, after a section dedicated to the importance of quantifying liver steatosis in patients with non-alcoholic fatty liver disease and another section dedicated to the assessment of liver fat with magnetic resonance, the current clinical studies performed using QUS are summarized. These new methods include spectral-based techniques and techniques based on envelope statistics. The spectral-based techniques that have been used in clinical studies are those estimating the attenuation coefficient and those estimating the backscatter coefficient. Clinical studies that have used tools based on the envelope statistics of the backscattered ultrasound are those performed by using the acoustic structure quantification or other parameters derived from it, such as the normalized local variance, and that performed by estimating the speed of sound. Experts' opinions are reported.

In this work, a novel calibration approach for minor and trace element quantification in LA-ICP-MS imaging of biological tissues is presented. Droplets of aqueous standard solutions are deposited onto pre-cut pieces of filter paper, allowed to dry, and sputtered with a thin gold layer for use as pseudo-internal standard. Analysis of the standards using LA-ICP-MS is performed using radial line-scans across the filters. In contrast to conventionally used preparation of matrix-matched tissue standards, the dried-droplet approach offers a variety of advantages: The standards are easy to prepare, no characterization of the standards using acid digestion is required, no handling of biological materials is necessary, and the concentration range, as well the number of investigated analytes is almost unlimited. The proposed quantification method has been verified using homogenized tissue standards with known analyte concentrations before being applied to a human malignant mesothelioma biopsy from a patient who had not received any chemotherapeutic treatment. Elemental distribution images were acquired at a lateral resolution of 40 μm per pixel, limits of detection ranging from 0.1 μg g(-1) (Mn, Ni, Cu, Zn) to 13.2 μg g(-1) (K) were reached.

[要旨]肝臓は, 脂質や糖質代謝の中心であり, 肥満に伴うメタボリックシンドロームの進展にも深くかかわる. 肥満に伴う肝臓の生化学的・病理学的な変化を包括的にとらえる手法を確立できれば, 生活習慣病を予防する薬剤の開発や, 生活習慣病予防効果を持つ食品機能性成分の分子機構の解明等に寄与すると考えられる. そこで, 本研究では, 組織中のタンパク質を網羅的に定量可能なSWATH質量分析法を用い, 高脂肪食誘導肥満マウスの肝臓におけるタンパク質発現プロファイルを解析し, 肥満に伴う肝臓の生化学的変化を捉えることができるかどうかを検討した. 6週齢のC57/BL/6J雄マウスを0.1%のセルロースを含有した高脂肪食（HD群, n=6）, および普通食（ND群, n=5）で5週間飼育し, 16時間の絶食後に肝臓を採取した. 肝臓タンパク質をトリプシン処理し, 生じたペプチドをnanoLC-MS/MS （AB sciex社TripleTOF5600+）で解析した. DDA法で得たMS/MSデータをUniprot Mouse databaseに対して検索し, Ion libraryを作成した. SWATH測定ではQ1 window幅を25 Daに固定し, m/z = 400 – 1000の範囲で25 windowを設定した. 各MS/MS測定の測定m/z範囲は100 – 1500 とし, 積算時間は100 msecとした. DDA測定によって, 1249個のタンパク質（10880個のペプチド）が同定された. このうち, SWATH法により768個のタンパク質が定量可能であった. これらのタンパク質には, 各種代謝酵素, 解毒酵素など肝臓を特徴づけるタンパク質が多数確認された. 得られた肝臓タンパク質プロファイルを主成分分析（PCA）, および主成分判別分析（PC-DA）にかけたところ, ND群とHD群は明確に分離され, それぞれ特徴的なタンパク質プロファイルを有していることが示された. ND群に比べて, HD群では解毒や呼吸鎖に関わる多くの酵素が減少しており, このうち一部は以前の報告と一致していた. また, HD群では細胞自律的免疫に関わるタンパク質や脂質代謝に関わる酵素が増加していた. 脂質代謝に関わる酵素には, ステロール代謝に関わる酵素が多く含まれ, 肥満に伴うコレステロールの増加を反映していると考えられた. [Abstract]Liver is a center of carbohydrate and lipid metabolism and deeply involved with the development of metabolic syndrome. If a method to comprehensively capture the biological and pathological changes in the liver caused by obesity could be established, it would contribute to the development of drugs to prevent lifestyle-related diseases, or to the elucidation of the molecular mechanism of food functional ingredients having life-style related disease prevention effect. In this study, we performed SWATH-mass spectrometry of livers obtained from high-fat diet （HD）-induced obese mice. Six weeks old C57/BL/6J male mice were fed with HD containing 0.1% cellulose, or normal diet （ND） for 5 weeks. On the day 35, livers were collected after 16 h of fasting. Proteins extracted from the liver-tissues were digested by trypsin, and the resulting peptides （3 ug） were analyzed by nanoLC-MS/MS （AB sciex, TripleTOF5600+ system）. Ion library of liver protein digests was built by data dependent analyses （DDA） and database search against Uniprot mouse protein database. In SWATH analysis, Q1 window range was fixed at 25 Da （24 windows along with m/z = 400 -1000）. Mass range, and accumulation time of MS/MS were set at 100 - 1500 and 100 msec, respectively. By the DDA analyses and database-search, 1249 proteins （10880 peptides） were identified. Among the identified proteins, 768 proteins were successfully quantified by the SWATH experiments. The quantified proteins containing those involved in carbohydrate metabolism, lipid metabolism, detoxification, and so on. PCA and PC-DA analysis of the liver proteome profiles clearly separated the HD and ND groups, indicating that the two groups had the distinctive proteome profiles. Compared to the ND, the amount of many proteins involved in detoxification and respiratory chain decreased in the HFD group, some of which were consistent with previous reports, while the amount of proteins involved in cell-autonomous immunity or lipid metabolism increased in the HFD group. Especially, many steroid-metabolizing enzymes were increased, which was thought to reflect the obesity-induced increase in liver or serum cholesterol.

Bioactive substances in the Biobio River Basin in Chile were examined by deploying Semi Permeable Membrane Devices (SPMDs) upstream and downstream of 4 pulp mill effluent discharges. Androgenic and estrogenic activity of SPMD extracts were then evaluated using in vitro fish sex steroid receptor binding assays. The results indicated the occurrence of estrogenic type compounds associated with one of the mill discharges. A significant correlation among the presence of these compounds, an increase in gonadosomatic index GSI and induction of hepatic EROD activity of two native fish species was observed. However, no significant presence of mature oocytes in female gonads was detected. Although EROD induction was observed in sites impacted by mill effluents, an increase of its activity occurred towards the downstream areas, suggesting other non-mill sources. More research is needed to understand the environmental changes in context of the new technological improvements in treatment systems to MBBR (Moving Bed Biofilm Reactor) recently implemented by the pulp mill industries.

Pulp and paper mill effluents (PPMEs) have been shown to increase gonad size, cause early maturation, and disrupt hormone functions in native and non-native Chilean fish. In this study, we assessed reproductive (plasma vitellogenin; VTG, gonad development) and metabolic (ethoxyresorufin-. O-deethylase activity; EROD) end points, relative liver size (LSI) and condition factor (K) of juvenile female and male rainbow trout exposed to effluents. Unlike previous studies, which have focus either on the specific effects of effluent on fish in laboratory exposures or biotic population statuses downstream of discharge sites, we simultaneously assessed the impacts of PPMES on trout using two approaches: (1) laboratory exposures of tertiary treated PPME produced from processing Eucalyptus globulus or Pinus radiata; and (2) in situ bioassay downstream of the combined discharge of the same pulp mill. Despite an increase in the average gonadosomatic index (GSI) in exposed fish, no statistical differences in gonad size between exposed and unexposed individuals was detected. However, both female and male fish exposed to effluents showed significantly higher concentrations of plasma VTG, so more in fish exposed to Eucalyptus-based effluent when compared to Pinus PPME. In addition, male fish showed intersex characteristics in all exposure assays (Eucaliptus and Pinus) and, despite the low concentration of effluent in the river (

Valnemulin (VAL) and tiamulin (TIA) are pleuromutilin antibiotics used primarily for treating bacterial infections in swine or other food animals. Furthermore, VAL and TIA are also employed as feed additives to promote animal growth. However, the illegal use of VAL and TIA could cause a series of hazards to consumers. Here, VAL was designed to be conjugated with bovine serum protein to prepare immunogen. A highly sensitive monoclonal antibody that recognized both VAL and TIA has been successfully produced. Moreover, an immunochromatographic strip assay for rapidly screening VAL and TIA in porcine liver was established with visual detection limits (cutoff values) of 50 and 25 ng/g, respectively. The IC50 values calculated from the equation of the standard curve were 6.06 and 3.45 ng/g and the limits of detection were 0.96 and 0.29 ng/g for VAL and TIA. According to the recovery experiment results, the test strip exhibited acceptable accuracy and precision. Generally, the proposed strip provided a practical tool for the detection of VAL and TIA. PRACTICAL APPLICATION: We produced a highly sensitive monoclonal antibody and developed an immunoassay strip for simultaneously monitoring TIA and VAL. Additionally it was preliminarily confirmed that the rapid detection tool was suitable for screening TIA and VAL in porcine liver.

Children comprise one-fifth of Europe's population. Promoting child health and development is of key importance for society and its future. This position paper highlights opportunities of investing in gastrointestinal, liver, and nutritional research to promote child health and delineates priorities for research. Investing in child health plays a key role in the promotion of population health, well-being, and disease prevention lifelong, with large health economic benefits. Major opportunities for improving knowledge and translational application arise from recent scientific and technological developments, for example, the long-term impact of early environmental cues interacting with genes. Personalised approaches to therapy and prevention should be enhanced. Deciphering the microbiome and its effects on functions can help in promoting long-term health. Epigenetic research can help to understand how early environmental factors influence later gastrointestinal and hepatic health and disease. A linked nutrition and physical activity strategy can promote health and prevent nutritional deficiencies, inactivity, and chronic noncommunicable diseases, such as diabetes, to ensure optimal health and cognition. Special attention should be devoted to populations with low socioeconomic status, migrant background, and ethnic minorities, and to critical life periods, including pregnancy, lactation, infancy, and childhood. Improved understanding of optimal nutrition and on maintaining gut and liver homeostasis throughout childhood will help prevent chronic diseases in later life.

Increased ethoxyresorufin-O-deethylase (EROD) activity is a sensitive biomarker of exposure to the chemicals which activate the aryl hydrocarbon receptor (AhR) and induce the cytochrome P450 system, such as many polychlorinated dibenzo-p-dioxins (PCDDs), polychlorinated dibenzofurans (PCDFs) and polychlorinated biphenyls (PCBs). Pulp bleaching was one of the main sources of PCDDs and PCDFs until elemental chlorine free (ECF) and total chlorine free bleaching processes since 1990s have remarkably decreased but not completely eliminate discharges of these chemicals. In addition, historically contaminated sediments may act as a source of these persistent contaminants. In this study, the contamination history and recovery of a watercourse heavily loaded by the chemical wood industry were studied by analyzing PCDDs, PCDFs and PCBs from vertical sediment samples and by measuring hepatic EROD activity from rainbow trout intraperitoneally dosed with the sediment extracts. No PCDDs or PCDFs were found above the chromatographic limit of detection from the study area and only small amounts of PCB congeners 101, 138, 153, and 180 were present. No increased EROD activity was observed in fish indicating the absence of any AhR-activating compounds in the surface sediment, to about 15 cm depth, representing about the last 20 years when kraft pulping and ECF bleaching with activated wastewater treatment have been used. It can be concluded that nowadays organochlorines and other AhR-ligands do not harm the previously heavily polluted watercourse.

In recent years, there has been growing interest in the possible use of nutraceuticals to improve and optimize dyslipidemia control and therapy. Based on the data from available studies, nutraceuticals might help patients obtain theraputic lipid goals and reduce cardiovascular residual risk. Some nutraceuticals have essential lipid-lowering properties confirmed in studies; some might also have possible positive effects on nonlipid cardiovascular risk factors and have been shown to improve early markers of vascular health such as endothelial function and pulse wave velocity. However, the clinical evidence supporting the use of a single lipid-lowering nutraceutical or a combination of them is largely variable and, for many of the nutraceuticals, the evidence is very limited and, therefore, often debatable. The purpose of this position paper is to provide consensus-based recommendations for the optimal use of lipid-lowering nutraceuticals to manage dyslipidemia in patients who are still not on statin therapy, patients who are on statin or combination therapy but have not achieved lipid goals, and patients with statin intolerance. This statement is intended for physicians and other healthcare professionals engaged in the diagnosis and management of patients with lipid disorders, especially in the primary care setting.

Background:Clinical presentations of Wilson's disease (WD) in childhood ranges from asymptomatic liver disease to cirrhosis or acute liver failure, whereas neurological and psychiatric symptoms are rare. The basic diagnostic approach includes serum ceruloplasmin and 24-hour urinary copper excretion. Final diagnosis of WD can be established using a diagnostic scoring system based on symptoms, biochemical tests assessing copper metabolism, and molecular analysis of mutations in the ATP7B gene. Pharmacological treatment is life-long and aims at removal of copper excess by chelating agents as D-penicillamine, trientine, or inhibition of intestinal copper absorption with zinc salts. Acute liver failure often requires liver transplantation. This publication aims to provide recommendations for diagnosis, treatment, and follow-up of WD in children.Methods:Questions addressing the diagnosis, treatment, and follow-up of WD in children were formulated by a core group of ESPGHAN members. A systematic literature search on WD using MEDLINE, EMBASE, Cochrane Database from 1990 to 2016 was performed focusing on prospective and retrospective studies in children. Quality of evidence was assessed according to the GRADE system. Expert opinion supported recommendations where the evidence was regarded as weak. The ESPGHAN core group and ESPGHAN Hepatology Committee members voted on each recommendation, using the nominal voting technique.

Diacetyl (2,3-butanedione) is a yellowish liquid that is usually mixed with other ingredients to produce butter flavor or other flavors in a variety of food products. Inhalation of butter flavoring vapors was first associated with clinical bronchiolitis obliterans among workers in microwave popcorn production. Recent findings have shown irreversible obstructive lung disease among workers not only in the microwave popcorn industry, but also in flavoring manufacture, and in chemical synthesis of diacetyl, a predominant chemical for butter flavoring. It has been reported that perfluorochemicals utilized in food packaging are migrating into foods and may be sources of oral exposure. Relatively small quantities of perfluorochemicals are used in the manufacturing of paper or paperboard that is in direct contact with food to repel oil or grease and water. Because of recent concerns about perfluorochemicals such as those found on microwave popcorn bags (e.g. Lodyne P208E®) and diacetyl in foods, we evaluated both compounds for mutagenicity using the mammalian cell gene mutation assay in L5178Y mouse lymphoma cells. Lodyne P208E® was less toxic than diacetyl and did not induce a mutagenic response. Diacetyl induced a highly mutagenic response in the L5178Y mouse lymphoma mutation assay in the presence of human liver S9 for activation. The increase in the frequency of small colonies in the assay with diacetyl indicates that diacetyl causes damage to multiple loci on chromosome 11 in addition to functional loss of the thymidine kinase locus.

Female mosquitofish exposed to pulp and paper mill effluent (PME) in the Fenholloway River, Florida, USA have masculinized secondary sex characteristics and altered aromatase enzyme activity. We and others have shown that the Fenholloway River PME contains androgenic and progestogenic substance(s). The present study was designed to test the hypothesis that the development and reproductive health of PME-exposed Fenholloway River mosquitofish are altered compared to mosquitofish living in Econfina River, which is the reference site. Fish were collected on a single day from both sites in June and August 1999 and January and June 2000. We compared standard length, anal fin length and segment number; body, liver, and gonad mass; and number of eggs and embryos from Fenholloway and Econfina River mosquitofish. The data were analyzed collectively for generalized site effect, for site effects during reproductive and nonreproductive seasons, and for repeatability of site effects between years. Mosquitofish exposed to PME in the Fenholloway River were generally smaller in length and mass, anal fin segment number was greater, and the number of embryos, but not oocytes, was significantly decreased compared to the reference site fish. Anal fin length and segment number and liver and testis masses were generally greater in Fenholloway compared to the Econfina River males. The importance of this study is that we have documented masculinized development and decreased embryo production in PME-exposed mosquitofish and that these site effects are generally consistent across seasons and between years.

We report here on studies designed to asses the effects of paper mill effluents on non-reproductive functions of free-ranging and captive Florida largemouth bass (Micropterus salmoides floridanus). This was accomplished by conducting an outdoor tank study, in which fish were exposed to well water or to 10%, 20%, 40%, and 80% full strength effluent for 28 or 56 days, and by sampling largemouth bass from sites within the St. Johns River, Florida, upstream and downstream from a paper mill plant. Blood and plasma samples from fish from the tank study and from fish sampled from the ambient sites were analyzed for over 20 variables. We also determined liver and spleen weights and examined them histologically. The most significant finding from the tank study was an increase in the concentration of albumin and hepatosomatic index for bass exposed toor = 20% effluents for 56 days. Spleenosomatic index and number of melanomacrophage centers were decreased in bass from effluent-dominated sites (Palatka and Rice Creek), whereas concentrations of calcium, phosphorous, glucose, and creatinine were elevated in fish from these sites, compared to fish from reference streams. Fish from Rice Creek also had fewer red blood cells, and male bass from Palatka had lower concentrations of cholesterol. Plasma concentrations of albumin and hepatic concentrations of glutathione were elevated in males from Palatka, and both females and males from Rice Creek had higher concentrations of globulin. These results indicate a complex pattern of effects of paper mill effluents on several physiological functions. However, despite the myriad of treatment and site-related effects, most physiological parameters fell within normal ranges when compared to reports on largemouth bass and other freshwater species.

While recent evidence indicates that the generation of reactive oxygen species (ROS) and associated oxidative damage are frequently observed in fish populations with exposure to pulp and paper mill effluents, the potential for ROS generation from municipal sewage effluents has not been addressed. This study investigates the utility of measures of oxidative stress in delineating the effects of both pulp and paper mill and municipal sewage discharges. Longnose sucker (Catostomus catostomus) were collected below three pulp and paper mill and two municipal sewage effluent discharges over a 3-year period within the Wapiti and Athabasca River systems in northern Alberta. Biochemical responses in longnose sucker varied between the two rivers systems, with more pronounced changes occurring within the Wapiti River. Of the suite of biochemical parameters examined, fatty acyl-CoA oxidase (FAO) activity was the most sensitive indicator of pulp and paper mill exposure, but was only infrequently induced with exposure to municipal sewage effluent. Hepatic and gonadal 2-thiobarbituric acid reactive substances, lipid hydroperoxides, and hepatic free iron were less consistently elevated with exposure to pulp and paper mill effluent than FAO activity, and were also only infrequently altered with sewage effluent exposure. Hepatic ascorbic acid, liver somatic index, and condition factor were consistently altered with exposure to both sewage and pulp and paper mill effluents. While specific biochemical and organismal responses varied with effluents and time, the collective suite of oxidative stress endpoints proved to be useful tools in identifying relative influences of municipal sewage and pulp and paper mill effluent on fish populations in adjacent receiving waters.

Laboratory tests have been used to assess the regulatory and research questions related to the effects of pulp mill effluents on aquatic biota. Acute, short-term laboratory tests have clearly shown the improvement in final effluent quality following installation of secondary treatment at Canadian pulp mills. In an effort to predict and investigate impacts on wild fish, laboratory bioassays were developed to examine sublethal endpoints: induction of hepatic mixed function oxygenase activity and reduction of sex steroid concentrations. These laboratory assays have been used to assess whole effluents, specific chemicals, and components of pulp mill processes, and to discriminate between historical and present-day effluent discharges. These tests have shown that induction of mixed-function oxygenase activity and reduction of sex steroid concentrations are produced by effluents from a variety of mill types, with and without chlorine bleaching, in hardwood and softwood pulping facilities, and before and after effluent treatment. These short-term bioassays have enabled reductions in sex steroid concentrations to be linked to mill process streams, and have provided information on effective waste stream treatment. Longer term, life-cycle fish bioassays have shown that chronic exposure to pulp mill effluents commonly results in growth enhancement, liver enlargement, and decreases in gonad size, secondary sex characteristics, and fecundity. These long-term laboratory exposures are able to mimic the most commonly observed alterations of wild fish exposed to pulp mill effluents: increases in condition factors, increases in liver-somatic indices, and decreases in gonadosomatic indices. This pattern of response is a combination of nutrient enrichment with metabolic disruption. The most sensitive and biologically meaningful endpoint is decreased reproduction in fish life-cycle exposures. As the laboratory tests move forward into the next decade, attention will focus on the reproductive endpoints and on the possibility of shortening the fish bioassays while still maintaining sensitivity and biological relevance.

Several environmental contaminants have been found in paper mill effluent including substances with androgenic properties. The presence of androgenic substances in paper mill effluent was also indicated in the present study which demonstrated masculinized anal fins of female mosquitofish from the paper mill effluent contaminated Fenholloway River. In addition, when compared to the nearby Econfina River, which does not receive paper mill effluent, fewer females from the contaminated river were pregnant, they were smaller and their estradiol concentration exhibited greater variation. Males from the Fenholloway River and the Econfina River had similar sperm counts, but the testes were larger and greater variation in testosterone concentration was observed in male fish from the Fenholloway River. In males and females from the Fenholloway River, liver weights were increased and computer-aided behavior analysis demonstrated a reduction in their social behavior when compared to reference fish. We conclude that a number of sexual characteristics were affected in mosquitofish living in the paper mill contaminated Fenholloway River, with possible adverse effects on the reproduction of this population.

Item does not contain fulltext We recommend that intestinal failure associated liver disease (IFALD) should be diagnosed by the presence of abnormal liver function tests and/or evidence of radiological and/or histological liver abnormalities occurring in an individual with IF, in the absence of another primary parenchymal liver pathology (e.g. viral or autoimmune hepatitis), other hepatotoxic factors (e.g. alcohol/medication) or biliary obstruction. The presence or absence of sepsis should be noted, along with the duration of PN administration. Abnormal liver histology is not mandatory for a diagnosis of IFALD and the decision to perform a liver biopsy should be made on a case-by-case basis, but should be particularly considered in those with a persistent abnormal conjugated bilirubin in the absence of intra or extra-hepatic cholestasis on radiological imaging and/or persistent or worsening hyperbilirubinaemia despite resolution of any underlying sepsis and/or any clinical or radiological features of chronic liver disease. Nutritional approaches aimed at minimising PN overfeeding and optimising oral/enteral nutrition should be instituted to prevent and/or manage IFALD. We further recommend that the lipid administered is limited to less than 1 g/kg/day, and the prescribed omega-6/omega-3 PUFA ratio is reduced wherever possible. For patients with any evidence of progressive hepatic fibrosis or overt liver failure, combined intestinal and liver transplantation should be considered.

Clinical presentations of Wilson's disease (WD) in childhood ranges from asymptomatic liver disease to cirrhosis or acute liver failure, whereas neurological and psychiatric symptoms are rare. The basic diagnostic approach includes serum ceruloplasmin and 24-hour urinary copper excretion. Final diagnosis of WD can be established using a diagnostic scoring system based on symptoms, biochemical tests assessing copper metabolism, and molecular analysis of mutations in the ATP7B gene. Pharmacological treatment is life-long and aims at removal of copper excess by chelating agents as D-penicillamine, trientine, or inhibition of intestinal copper absorption with zinc salts. Acute liver failure often requires liver transplantation. This publication aims to provide recommendations for diagnosis, treatment, and follow-up of WD in children.

Liver biopsy (LB) is still the criterion standard procedure for obtaining liver tissue for histopathological examination and a valuable tool in the diagnosis, prognosis, and management of many parenchymal liver diseases. The aim of this position paper is to summarise the present practice of paediatric LB and make recommendations about its performance. Although histological evaluation of the liver is important in assessing prognosis and exploring treatment, noninvasive techniques (ie, imaging, laboratory markers) may replace use of liver histology. The indications for LB are changing as present knowledge of aetiologies, pathomechanism, and therapeutic options in paediatric liver disease is evolving. Adult and paediatric literature was reviewed to assess the existing clinical practice of LB with focus on the technique, indications, risk of complications, and contraindications in paediatrics. This position paper presents types of LB, indications, complications, contraindications, and an essential checklist for paediatric LB.

Non-alcoholic fatty liver disease (NAFLD) is a highly prevalent chronic liver disease, and patient susceptibility to its onset and progression is influenced by several factors. In this study, we investigated whether altered hepatic DNA methylation in liver tissue correlates with the degree of severity of NAFLD-like liver injury induced by a high-fat and high-sucrose (HF/HS) diet in Collaborative Cross (CC) mice. Using genome-wide targeted bisulphite DNA methylation next-generation sequencing, we found that mice with different non-alcoholic fatty liver (NAFL) phenotypes could be distinguished by changes in hepatic DNA methylation profiles. Specifically, NAFL-prone male CC042 mice exhibited more prominent DNA methylation changes compared with male CC011 mice and female CC011 and CC042 mice that developed only a mild NAFL phenotype. Moreover, these mouse strains demonstrated different patterns of DNA methylation. While the HF/HS diet induced both DNA hypomethylation and DNA hypermethylation changes in all the mouse strains, the NAFL-prone male CC042 mice demonstrated a global predominance of DNA hypermethylation, whereas a more pronounced DNA hypomethylation pattern developed in the mild-NAFL phenotypic mice. In a targeted analysis of selected genes that contain differentially methylated regions (DMRs), we identified NAFL phenotype-associated differences in DNA methylation and gene expression of the Apoa4, Gls2, and Apom genes in severe NAFL-prone mice but not in mice with mild NAFL phenotypes. These changes in the expression of Apoa4 and Gls2 coincided with similar findings in a human in vitro cell model of diet-induced steatosis and in patients with NAFL. These results suggest that changes in the expression and DNA methylation status of these three genes may serve as a set of predictive markers for the development of NAFLD.

As a common cause of liver injury, hepatic ischemia/reperfusion injury (HIRI) happens in various clinical conditions including trauma, hepatectomy and liver transplantation. Since transcription factor Yin Yang 1 (YY1) was reported to be downregulated after ischemia/reperfusion (I/R) injury, we focused on YY1 to explore its function in HIRI by functional assays like Cell Counting Kit-8 (CCK-8) assays and flow cytometry assays. The RT-qPCR assay revealed that YY1 was downregulated in hepatocytes after I/R injury. The function assays disclosed that YY1 facilitated cell viability and proliferation, but hindered cell apoptosis in hepatocytes after I/R injury. Through mechanism assays including luciferase reporter assay, RIP and RNA pulldown assay, miR-186-5p was found to bind with YY1 and promote hepatocyte apoptosis by targeting YY1. Subsequently, we verified that small nucleolar RNA host gene 1 (SNHG1) could sponge miR-186-5p to upregulate YY1. Importantly, we figured out that YY1 had a positive regulation on SNHG1. Along the way, YY1 was identified as the upstream transcription factor for SNHG1. In conclusion, our study unveiled a novel competing endogenous RNA (ceRNA) pattern of SNHG1/miR-186-5p/YY1 positive feedback loop in hepatic I/R injury, which might provide new insight into prevention of HIRI during liver transplantation or hepatic surgery.

Mutations in the mitochondrial genome (mtDNA) are ubiquitous in humans and can lead to a broad spectrum of disorders. However, due to the presence of multiple mtDNA molecules in the cell, co-existence of mutant and wild-type mtDNAs (termed heteroplasmy) can mask disease phenotype unless a threshold of mutant molecules is reached. Importantly, the mutant mtDNA level can change across lifespan as mtDNA segregates in an allele- and cell-specific fashion, potentially leading to disease. Segregation of mtDNA is mainly evident in hepatic cells, resulting in an age-dependent increase of mtDNA variants, including non-synonymous potentially deleterious mutations. Here we modeled mtDNA segregation using a well-established heteroplasmic mouse line with mtDNA of NZB/BINJ and C57BL/6N origin on a C57BL/6N nuclear background. This mouse line showed a pronounced age-dependent NZB mtDNA accumulation in the liver, thus leading to enhanced respiration capacity per mtDNA molecule. Remarkably, liver-specific atg7 (autophagy related 7) knockout abolished NZB mtDNA accumulat ion, resulting in close-to-neutral mtDNA segregation through development into adulthood. prkn (parkin RBR E3 ubiquitin protein ligase) knockout also partially prevented NZB mtDNA accumulation in the liver, but to a lesser extent. Hence, we propose that age-related liver mtDNA segregation is a consequence of macroautophagic clearance of the less-fit mtDNA. Considering that NZB/BINJ and C57BL/6N mtDNAs have a level of divergence comparable to that between human Eurasian and African mtDNAs, these findings have potential implications for humans, including the safe use of mitochondrial replacement therapy.Abbreviations: Apob: apolipoprotein B; Atg1: autophagy-related 1; Atg7: autophagy related 7; Atp5a1: ATP synthase, H+ transporting, mitochondrial F1 complex, alpha subunit 1; BL6: C57BL/6N mouse strain; BNIP3: BCL2/adenovirus E1B interacting protein 3; FCCP: carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MAP1LC3A: microtubule-associated protein 1 light chain 3 alpha; MAP1LC3B: microtubule-associated protein 1 light chain 3 beta; mt-Atp8: mitochondrially encoded ATP synthase 8; MT-CO1: mitochondrially encoded cytochrome c oxidase I; MT-CO2: mitochondrially encoded cytochrome c oxidase II; mt-Co3: mitochondrially encoded cytochrome c oxidase III; mt-Cytb: mitochondrially encoded cytochrome b; mtDNA: mitochondrial DNA; MUL1: mitochondrial ubiquitin ligase activator of NFKB 1; nDNA: nuclear DNA; Ndufa9: NADH:ubiquinone oxireductase subunit A9; NDUFB8: NADH:ubiquinone oxireductase subunit B8; Nnt: nicotinamide nucleotide transhydrogenase; NZB: NZB/BINJ mouse strain; OXPHOS: oxidative phosphorylation; PINK1: PTEN induced putative kinase 1; Polg2: polymerase (DNA directed), gamma 2, accessory subunit; Ppara: peroxisome proliferator activated receptor alpha; Ppia: peptidylprolyl isomerase A; Prkn: parkin RBR E3 ubiquitin protein ligase; P10: post-natal day 10; P21: post-natal day 21; P100: post-natal day 100; qPCR: quantitative polymerase chain reaction; Rpl19: ribosomal protein L19; Rps18: ribosomal protein S18; SD: standard deviation; SEM: standard error of the mean; SDHB: succinate dehydrogenase complex, subunit B, iron sulfur (Ip); SQSTM1: sequestosome 1; Ssbp1: single-stranded DNA binding protein 1; TFAM: transcription factor A, mitochondrial; Tfb1m: transcription factor B1, mitochondrial; Tfb2m: transcription factor B2, mitochondrial; TOMM20: translocase of outer mitochondrial membrane 20; UQCRC2: ubiquinol cytochrome c reductase core protein 2; WT: wild-type.

To develop an effective hybrid bioartificial liver (BAL) device, the material of the scaffold is very important to support hepatocytes that have both growth ability and hepatic differentiated functions. In this study we used paper (Kimwipe, Kimberly-Clark Corp., Roswell, GA, U.S.A.) as a scaffold. Primary hepatocytes isolated from a normal adult rat liver could proliferate on the paper. The secretion of albumin into culture medium by the cells on the paper increased with time in culture and, compared to the cells on dishes, the amount of 48 h albumin secretion at Day 10 was two times larger. Perpendicular sections of hepatocytes on the paper revealed that the cells fell into cavities made by intersecting fibers, piled up, and formed three to four layers. The piled-up cells changed their shape from flat to cuboidal and enlarged their cytoplasm, which was rich in organelles such as mitochondria and peroxisomes with a nucleoid. In addition, they formed bile canalicular structures between the cells. Their morphological appearance was similar to in vivo hepatocytes. Paper (Kimwipe) may be a good candidate as a scaffold to make a BAL device.

Juvenile whitefish (Coregonus lavaretus L. s.l.) were exposed by caging in the field to diluted effluents from three operating pulp, paper, and paperboard mills in Southern Lake Saimaa, Finland. The expression of the vitellogenin gene, used as a biomarker of estrogenic contamination of effluents, was measured using a Northern blotting method. Increased mRNA levels, the most specific and reliable evidence for estrogen receptor-mediated actions in vivo, were found in fish caged in the vicinity of one of three mills studied. This mill was found to discharge wood-derived compounds, such as sterols and resin acids, into Lake Saimaa in amounts considerably exceeding those from the other two mills. The increased vitellogenin gene expression suggests that the effluent is a source of estrogenic contaminants.

Autoimmune hepatitis (AIH) is an immune-mediated chronic inflammatory liver disease, and its pathogenesis is not fully understood. Our previous study discovered that receptor interacting protein kinase 3 (RIP3) is correlated with serum transaminase levels in AIH patients. However, its role and underlying mechanism in AIH are poorly understood. Here, we detected the increased expression and activation of RIP3 in livers of patients and animal models with AIH. The inhibition of RIP3 kinase by GSK872 prevented concanavalin A (ConA)-induced immune-mediated hepatitis (IMH) by reduced hepatic proinflammatory cytokines and immune cells including Th17 cells and macrophages. Further experiments revealed that RIP3 inhibition resulted in an increase in CD11b+Gr1+ myeloid-derived suppressor cells (MDSCs) with immunoregulatory properties in the liver, spleen, and peripheral blood. Moreover, the depletion of Gr-1+ MDSCs abrogated the protective effect and immune suppression function of GSK872 in ConA-induced IMH. Altogether, our data demonstrate that RIP3 blockade prevents ConA-induced IMH through promoting MDSCs infiltration. Inhibition of RIP3 kinase may be a novel therapeutic avenue for AIH treatment.

Assessment and enhancement of liver clearance are promising strategies for protection of liver from various liver diseases. Iron-tannic nanoparticles (FTs) were previously considered as imageable autophagic enhancers with biodegradation potential. Herein, we present a new approach for utilizing Iron-tannic nanoparticles (FTs) as a tool for imaging and increasing liver clearance. Pharmacokinetic profiling suggested that FTs were initially found in blood circulation and thereafter were distributed to the liver. By using MR imaging (T1 weighted), maximum MRI signal enhancement was found to occur after 30 minutes post-injection (i.v.) and gradually decreased afterward. Decreasing MRI signal may be due to FTs metabolism by the liver. By assessing imaging-derived pharmacokinetics, we can simply determine the rate constant of liver degradation of FTs. Potentially, we might use this parameter to monitor liver function, where its clearance is of concern. Once functional implication of FTs in liver clearance was investigated, FTs were found to induce hepatocyte autophagy along with activation of lysosomes. Consequently, the hepatocytes were capable of efficiently clearing cellular debris. From these results, it is clear that FTs should be considered as a molecular tool for quantitative MRI-derived liver function assessment, and for enhancing clearance function in liver parenchyma. Hopefully, our findings will pave the way to develop new strategies for non-invasive assessment and enhancement of liver clearance.

[cat] En el primer estudi confirmem que el tractament amb enoxaparina es capaç de disminuir la pressió portal en rates amb cirrosi establerta i hipertensió portal. A més, la millora en la pressió portal es va produir sense efectes en el flux sanguini portal, suggerint una disminució en la resistència vascular intrahepàtica. La falta d’efectes de l’administració aguda d’enoxaparina en la pressió portal, en els paràmetres hemodinàmics sistèmics, i en el contingut de superòxid o GMPc, suggereix que l’enoxaparina no té un efecte directe sobre el component dinàmic de la resistència vascular intrahepàtica. Per acabar de confirmar aquests resultats, es va examinar la resposta dependent d’endoteli al vasodilatador acetilcolina i no es van observar diferències entre les rates tractades amb enoxaparina i les rates tractades amb vehicle en cap dels dos models de cirrosi.. A més a més, tot i que en els tractaments de més llarga durada amb enoxaparina es van observar sobretot millores en les alteracions arquitecturals, es van observar també canvis en el to vascular. Es van disminuir els nivells d’estrès oxidatiu, de proteïnes nitrotirosinades i es va augmentar en el contingut de GMPc. A més a més, es va observar una disminució de la ratio p-Moesina/Moesina en les rates tractades amb enoxaparina, suggerint que el tractament amb enoxaparina pot modular el to vascular mitjançant la relaxació de les CHE, almenys a llarg termini. Tot i això, s’ha de tenir en compte que aquest canvis en el to vascular poden ser deguts a la millora de l’alteració estructural observada. És cert que el tractament amb enoxaparina ja es va descriure com a un bon tractament per a millorar la fibrosi hepàtica en models de dany hepàtic lleu. El nostre estudi amplia aquestes observacions a models amb cirrosi establerta. El tractament amb enoxaparina va ser iniciat en el moment on es va aturar l’administració del tòxic (tant CCl4 com TAA) com a model de regressió de malaltia avançada, o es va administrar alhora amb el tòxic, com a model de prevenció de la progressió de la cirrosi, en el model de TAA. Això ens va permetre analitzar els efectes del tractament amb enoxaparina en dos escenaris diferents. En el model de regressió amb CCl4, les rates que van rebre durant 3 setmanes el tractament van tenir més temps de regressió espontània de la fibrosi que les rates que van rebre només una setmana de tractament. Tot i això, tant les d’una setmana de tractament com les de 3 setmanes de tractament van tenir una disminució del 25% del contingut de col·lagen analitzat per tinció de Sirius Red. Això també es va observar en el model de cirrosi de TAA tant en regressió com en prevenció. Tot els resultats van cap a la mateixa direcció i suggereixen el paper de l’enoxaparina en disminució de la fibrosi hepàtica. A més, es va observar una disminució dels nivells d’activació de les CHE en les rates tractades amb enoxaparina, tant a nivell de teixit com en cèl·lula aïllada de rata tractada. A part de l’efecte de l’enoxaparina en la fibrosi hepàtica, es va observar la influència de del tractament en la prevenció d’esdeveniments trombòtics en la microcirculació hepàtica. De fet, s’ha suggerit que aquests podrien jugar un paper important en la progressió de la fibrosi hepàtica. Per explorar aquest efecte hem avaluat el contingut de fibrina en el parènquima hepàtic com a marcador de la formació de microtrombes. Els fetges de rates tractades amb enoxaparina van tenir una menor deposició de fibrina. Aquest fet ja es va observar en les rates tractades durant una setmana, però l’efecte va ser més gran i significatiu en les rates tractades durant 3 setmanes. Aquestes troballes confirmen la connexió que hi ha entre trombosi i fibrosi hepàtica i consolida el concepte de l’ús dels anticoagulants per a la prevenció/dissolució dels microtrombes per a millorar la fibrosi hepàtica. Per contra, no es van observar canvis a nivell d’inflamació. Per tant, encara que no podem descartar completament el potencial efecte del tractament amb enoxaparina en la reducció de la inflamació, no sembla que aquests sigui el principal mecanisme responsable de la millora les alteracions arquitecturals en la cirrosi hepàtica. És important remarcar que els efectes beneficiosos del tractament amb enoxaparina en la reducció de la pressió portal, resistència vascular intrahepàtica i fibrosi hepàtica no es van associar a efectes deleteris en els animals. No es van observar diferencies en els enzims hepàtics, ni es va observar sagnats durant el tractament o durant l’estudi hemodinàmic. En el segon estudi es va avaluar l’efecte de l’anticoagulant oral, Rivaroxaban, en la cirrosi hepàtica. Com ja es va veure en el primer estudi, la teràpia anticoagulant pot ser una bona estratègia terapèutica per al tractament de la hipertensió portal en la cirrosi. Tot i això, les heparines de baix pes molecular són dependents dels nivells sanguinis d’antitrombina, que de fet poden estar disminuïts en els pacients cirròtics. Es per això que vam estudiar els efectes de Rivaroxaban, que és un inhibidor directe del factor Xa (tant el Factor Xa lliure com el que es troba unit al complex de protrombinasa) i és independent dels nivells d’antitrombina. A més a més, Rivaroxaban té l’avantatge de ser biodisponible via oral i té una acció ràpida. Tot això, fa de rivaroxaban un anticoagulant més previsible i potencialment més eficient comparat amb altres heparines. En aquest estudi hem confirmat els efectes beneficiosos de l’ús d’anticoagulants per a millora la hipertensió portal en la cirrosi. Hem demostrat que l’administració oral de rivaroxaban disminueix els nivells d’estrès oxidatiu, millora la biodisponibilitat de NO, desactiva les CHE i millora la hipertensió portal en dos models de cirrosi en rata. La disminució de la pressió portal observada en les rates tractades amb rivaroxaban no es va associar a canvis en el flux sanguini portal en cap dels dos models, suggerint que el principal efecte ve de la disminució en la resistència vascular intrahepàtica. De fet, es va observar una disminució de la resistència vascular intrahepàtica en fetges de rates tractades amb rivaroxaban quan es va avaluar en un sistema de perfusió ex vivo amb el flux controlat. L’estudi dels mecanismes de la millora en la funció microcirculatòria hepàtica van mostrar que rivaroxaban disminuïa els nivells d’estrès oxidatiu, probablement pel menor reclutament de cèl·lules inflamatòries. També augmentava els nivells de GMPc, millorava la resposta a dosis creixents d’acetilcolina i disminuïa l’expressió del marcador d’activació de CES, el factor von Willebrand. Tot això indicava que el tractament amb rivaroxaban té un efecte important en la millora de la disfunció endotelial en el fetge cirròtic. D’altra banda també es va estudiar l’efecte de rivaroxaban sobre les CHE. Es va observar que el tractament amb rivaroxaban disminuïa els marcadors d’activació i proliferació de les CHE que correlacionen amb la seva capacitat de síntesi de matriu extracel·lular. Aquest efecte no es va associar a canvis importants en el contingut de fibra en el fetge. Degut als canvis observats en els nivells d’activació de les CHE, es temptador especular que un tractament més llarg amb rivaroxaban s’associaria a una disminució en els nivells de fibrosi hepàtica. Això està recolzat pels efectes beneficiosos observats a sobre les alteracions del parènquima: estudis ultraestructurals realitzats mitjançant microscopia electrònica van confirmar que les rates tractades amb rivaroxaban tenien una menor formació de membrana basal en els sinusoides hepàtics i que els hepatòcits tenen més projeccions, suggerint una millor estructura del sinusoid hepàtic. Finalment, es va confirmar una disminució del contingut de fibrina en les rates tractades amb rivaroxaban, suggerint una disminució de la microtrombosi hepàtica. Els efectes beneficiosos deguts al tractament amb rivaroxaban no es van associar a efectes deleteris en l’hemodinàmica sistèmica ni en un augment de sagnats durant el tractament. El tercer estudi aporta nova informació, fins ara no coneguda, sobre el potencial dany de l’estès oxidatiu mitocondrial en les cèl·lules hepàtiques durant la cirrosi i sobre l’ús dels antioxidants dirigits a mitocondri com a estratègia terapèutica per a la hipertensió portal i la cirrosi. Tal i com s’ha explicat anteriorment, les CHE són les principals cèl·lules implicades en la síntesi de matriu extracel·lular que contribueix en l’augment de la resistència vascular intrahepàtica en la cirrosi. Tenint en compte que l’augment de l’estrès oxidatiu en la cirrosi és un dels principals mecanismes que activen les CHE i que els mitocondris són un dels principals productors d’espècies reactives d’oxigen, aquest estudi es va centrar en avaluar els efectes de l’antioxidant dirigit a mitocondri, mitoquinona en l’estrès oxidatiu, fenotip de les CHE, la fibrosi hepàtica i la hipertensió portal en models pre-clínics de cirrosi. Els resultats del present estudi mostren que el tractament amb mitoquinona disminueix l’estrès oxidatiu hepàtic, desactiva les CHE i disminueix la fibrosi hepàtica i la hipertensió portal en les rates cirròtiques. Estudis anteriors van demostrar el rol de l’estrès oxidatiu en la fisiopatologia de la cirrosi i la hipertensió portal. En el present estudi demostrem que el tractament amb mitoquinona va aconseguir disminuir els nivells d’estrès oxidatiu, com altres teràpies utilitzades com ara el tempol o el resveratrol, però amb la diferència que la mitoquinona té l’avantatge de ser administrada oralment. A més a més, les altres teràpies antioxidants no poden dirigir-se a l’interior del mitocondri i, per tant, no poden contrarestar l’estrès oxidatiu mitocondrial que pot resultar en dany i disfunció mitocondrial i endegar una cascada de conseqüències deletèries per a la cèl·lula com ara la inflamació o la mort cel·lular. El nostre estudi demostra que part de l’estrès oxidatiu observat en els fetges cirròtics prové del mitocondris dels hepatòcits i de les CHE. A més, demostrem que la mitoquinona es capaç de disminuir l’estrès oxidatiu mitocondrial en els principals tipus cel·lulars hepàtics. Aquest augment de l’estrès oxidatiu mitocondrial en la cirrosi pot ser causat, almenys en part, per la disminució en l’expressió i l’activitat de la isoforma mitocondrial de la superòxid dismutasa, que s’ha vist disminuïda en la cirrosi. És important destacar que el tractament amb mitoquinona és capaç de disminuir l’activació de les CHE de rata tant in vitro com in vivo. A més, la desactivació de les CHE es va confirmar tant en CHE aïllades de biòpsies humanes, en la línia cel·lular humana de CHE, LX2, i en el model ex vivo per a l’estudi de mostres de fetge humanes, els talls de precisió de fetges humans (hPCLS). Aquests efectes es van evidenciar quan es va veure que la proliferació de les CHE estava disminuïda però no la seva viabilitat. Per confirmar la capacitat terapèutica de la mitoquinona, es va avaluar els seus efectes en dos models de cirrosi. Les observacions in vivo van demostrar que el tractament amb mitoquinona disminuïa la pressió portal, sense canvis en el flux sanguini portal, suggerint una disminució en la resistència vascular intrahepàtica. A més, els animals tractats amb mitoquinona van tenir una disminució del contingut de fibra hepàtica en ambdós models. Suggerint que la disminució de la fibrosi hepàtica és el principal mecanisme amb el que la mitoquinona disminueix la pressió portal, ja que no es van trobar efectes importants de la mitoquinona sobre el to vascular hepàtic almenys durant el temps de tractament estudiat, i que altres antioxidants sí que han demostrat una millora de la disfunció endotelial. A més, es sap que els nivells de les espècies reactives d’oxigen estan directament lligades a la inflamació. Per això vam analitzar l’expressió dels marcadors CD68 i CD163 com a marcadors d’inflamació. En aquest estudi vam observar una disminució del nombre de cèl·lules positives per CD68, suggerint un efecte de la mitoquinona en la reducció de la inflamació hepàtica. Aquests resultats només es van observar en el model de CCl4 i no en el de TAA, suggerint que la reducció de la inflamació pot contribuir a la millora de la fibrosi hepàtica però no és el principal mecanisme. Les diferències observades entre els dos models de cirrosi, pot ser degut a les diferències intrínseques dels dos models. De fet, s’ha descrit que el model induït per CCl4 acumula més estrès oxidatiu que el model per TAA, fent així el model per CCl4 més susceptible per a una teràpia antioxidant que no el model de TAA. Tot i això, cal destacar que tots els beneficis observats pel tractament amb mitoquinona no es van associar a efectes deleteris en els animals cirròtics en cap dels dos models., [eng] The present doctoral thesis is focused in the development of new therapeutical strategies for the treatment of portal hypertension, one of the main complications of cirrhosis. Increase in portal pressure is directly related to the increased intrahepatic vascular resistance, mainly due to increased vascular tone and increased hepatic fibrosis. Therefore, the main objective of the present doctoral thesis was to study the use of the anticoagulants enoxaparin and rivaroxaban, and the mitochondrial-directed antioxidant mitoquinone and to evaluate their effects on intrahepatic vascular resistance and portal pressure in preclinical models of cirrhosis with portal hypertension. On the one hand, the results of the present doctoral thesis concluded that the anticoagulants, both the anti-thrombin like enoxaparin and the anti-factor Xa (thrombin-independent) rivaroxaban significantly improve portal pressure of animals with cirrhosis and portal hypertension mainly due to improved vascular tone and hepatic fibrosis. On the other hand, we observed that mitochondrial oxidative stress was increased in cirrhotic livers and hepatocytes and hepatic stellate cells were the main cells producing mitochondrial oxidative stress. Furthermore, we confirmed the beneficial effects of antioxidants on cirrhosis. The results concluded that the mitochondrial-directed antioxidant mitoquinone decreased significantly portal pressure and intrahepatic vascular resistance. Moreover, an important deactivation of the main cells producing collagen was observed both in rat and in human cells. These results suggest that anticoagulant and antioxidant treatment may be a potential therapeutical strategy for the treatment of portal hypertension in cirrhotic patients., [spa] Esta tesis estudia el efecto de diferentes estrategias terapéuticas para el tratamiento de la hipertensión portal en models preclínicos de cirrosis. Se han estudiado los efectos de dos anticoagulantes (enoxaparina y rivaroxaban) y el efecto de un antioxidante dirigido a mitocondria (mitoquinona). En el primer estudio, se evaluó el efecto del anticoagulante enoxaparina en modelos preclínicos de cirrosis. Se observó que el tratamiento agudo con enoxaparina no afectó los parámetros sistémicos ni hemodinámicos evaluados. Tampoco modificó los niveles de GMPc o el contenido de estrés oxidativo hepático. El tratamiento a corto plazo (1 semana) disminuyó de manera significativa la presión portal en ratas cirróticas sin modificar el flujo. También disminuyó de manera significativa los niveles de estrés oxidativo y aumentó la biodisponibilidad de óxido nítrico (NO). Además, el tratamiento disminuyó un 26% la fibra hepática. Estos cambios se asociaron a una mejora del fenotipo de las células estrelladas hepáticas (CEH). También se observó una mejor en la microtrombosis hepática. El tratamiento a largo plazo (3 semanas) también disminuyó de manera significativa la presión portal en los dos modelos de cirrosis preclínica, sin cambios en los otros parámetros hemodinámicos. En este caso, también se observó una disminución del estrés oxidativo, pero sin mejora en la biodisponibilidad de NO. El tratamiento con enoxaparina redujo un 35% el área de fibrosis, cambios también asociados a una mejora del fenotipo de las CHE. También disminuyó el contenido de fibrina, mejorando así la microtrombosis hepática. Finalmente, también se observaron efectos beneficiosos en el modelo de prevención en ratas cirróticas por tioacetamida. La presión portal fue menor, en las ratas tratadas con enoxaparina, aunque los cambios no fueron tan marcados como en el modelo de regresión. También se redujo el área de fibrosis hepática, se mejoró el fenotipo de las CHE y hubo una menor microtrombosis hepática. En el segundo estudio, el anticoagulante oral anti factor Xa Rivaroxaban, disminuyó significativamente la presión portal en los dos modelos de cirrosis preclínica. La disminución de la presión portal no se asoció a cambios ni en el flujo sanguíneo portal ni en la presión arterial media, sugiriendo una disminución en la resistencia vascular intrahepática. Este hecho se confirmó ex vivo, donde se observó una disminución significativa de la resistencia vascular intrahepática del 35% en un sistema de perfusión con el flujo controlado. El tratamiento durante dos semanas disminuyó los niveles de estrés oxidativo, el número de células de Kupffer y aumentó la biodisponibilidad del NO. Las ratas tratadas con rivaroxaban tuvieron una mejor respuesta, aunque no significativa, al vasodilatador acetilcolina. Para confirmar el efecto beneficioso de rivaroxaban sobre el endotelio sinusoidal hepático, se evaluó el fenotipo de las células endoteliales sinusoidales y se observó una disminución en el marcador de activación, el factor von Willebrand, y una menor presencia de membrana basal. El tratamiento con rivaroxaban disminuyó el número y la activación de las CHE tanto a nivel de tejido como a nivel de célula aislada. No se observaron cambios en la apoptosis sugiriendo que el menor número de CHE vendría de una menor proliferación y no de un aumento en la muerte celular. Finalmente se observó que el tratamiento disminuía el contenido de fibrina sugiriendo una disminución de la microtrombosis hepática y se confirmó que los efectos de rivaroxaban sobre las CHE no era directos sino a través de la inhibición de la cascada de coagulación e inhibiendo la activación de los receptores PAR. En el tercer y último estudio se evaluó el efecto del tratamiento con un antioxidante dirigido a mitocondria. En un inicio se confirmó que en la cirrosis existe un aumento significativo de estrés oxidativo mitocondrial y que las principales células productores de éste estrés eran las CHE y los hepatocitos. El tratamiento con mitoquinona in vitro disminuyó significativamente el nivel de estrés oxidativo en todas las estirpes celulares hepáticas estudiadas. Además, disminuyó el nivel de activación de las CHE tanto de rata como humanas. Esto se confirmó en cortes ultrafinos de biopsias humanas y en líneas celulares. A nivel in vivo se observó que mitoquinona produjo un descenso significativo del nivel de estrés oxidativo hepático, tanto a nivel mitocondrial como total. A demás se disminuyó el contenido de proteínas nitrotirosinadas y la expresión génica de un factor inducido por el estrés oxidativo, Hif1a. Las ratas tratadas con mitoquinona tuvieron una disminución significativa de la presión portal y no se asoció a cambios ni en el flujo sanguíneo portal ni en la presión arterial media, sugiriendo una disminución en la resistencia vascular intrahepática. Mitoquinona también disminuyó el grado de fibrosis hepática y el nivel de activación de las CHE. Finalmente, se observó que el tratamiento con mitoquinona disminuyó significativamente el nivel de inflamación hepática. Para finalizar, esta tesis doctoral concluye que tanto el tratamiento con anticoagulantes, como el tratamiento con antioxidantes dirigidos a mitocondria pueden ser una potencial vía de tratamiento para la hipertensión portal en pacientes cirróticos.

Physiological and biochemical biomarker responses were studied in juvenile whitefish (Coregonus lavaretus L. s.l.) exposed experimentally to effluent from the forest industry. The large study area (609 km2), Southern Lake Saimaa, in Southeast Finland, receives 330,000 m3 d-1 of biologically and 55,000 m3 d-1 of chemically treated effluents, discharged from two integrated elementary chlorine free (ECF) bleached kraft pulp and paper mills, from one ECF pulp mill, and from one mill producing unbleached pulp and cardboard. The assessment of exposure to effluent discharged from the mills was based on lake water chlorophenolics (CPs) and resin acids (RAs) measured in samples collected from the 22 experimental sites along the area. Despite the low levels of effluent constituents in the lake, they were still accumulated in detectable levels in fish bile, indicating an exposure to the bioactive compounds of effluents. In comparison to the reference area, a two- to four-fold increase in ethoxyresorufin O-deethylase (EROD) activity was observed in whitefish exposed in the vicinity (1-6 km) of all the mills. However, cytochrome P450 1A1 (CYP1A1) gene expression was increased in only one of the receiving areas, indicating higher sensitivity of the EROD activity in the present study. There were no statistically significant correlations between EROD activity and the ambient water concentrations of the CPs, the RAs, or effluent dilution expressed by water sodium concentration. Neither bile chlorophenolics nor bile resin acids showed a significant correlation with EROD. No significant changes in circulating reproductive steroids, 17beta-estradiol and testosterone, in juvenile whitefish were observed. The vitellogenin gene was expressed in the vicinity of the pulp mill discharging the most wood-derived compounds, i.e. resin acids and wood-sterols, including beta-sitosterol. No differences were observed in plasma immunoglobulin M, glucose, or lactate concentrations between the effluent sources.

Perfluorooctanoic acid (PFOA) can accumulate in the livers of humans and animals via the food chain, resulting into liver injury, which is closely related to intestinal flora dysbiosis. Gastrodin has been reported to have hepatoprotective effect. However, whether gastrodin can alleviate PFOA-induced liver injury via modulating gut microbiota remains unclear. Herein, a PFOA-induced liver injury model was established by gavage of PFOA (5 mg/kg body weight) in 2% Tween 80 solution once daily for 6 weeks in mice, and then gastrodin in saline (20 mg/kg body weight) was used once daily for 8 weeks to treat liver damage. The biochemical indexes associated with liver function, oxidative stress, and inflammatory factors were examined. Hematoxylin-eosin staining was used to determine the liver histopathological changes. Besides, 16S rRNA sequencing was used to analyze the difference of gut microbiota between the model and treatment groups. The results showed that gastrodin significantly improved the oxidative stress caused by PFOA. Intestinal flora analysis showed that gastrodin treatment significantly increased the relative abundance of probiotics, such as Lactobacillus, Bifidobacterium, and Bacteroides, while the harmful bacteria, including Desulfovibrio were decreased. Gastrodin treatment also significantly increased the level of short-chain fatty acids (SCFAs), such as butyric acid and isobutyric acid. Spearman correlation analysis showed that the composition changes of gut microbiota and SCFAs increase were both beneficial to alleviate the liver injury caused by PFOA. To sum up, gastrodin can effectively alleviate PFOA-induced liver injury through regulating gut microbiota composition.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors worldwide, and chemoprevention represents a feasible treatment to reduce the mortality of this carcinoma. Mulberry fruit polysaccharides (MFP) possess immunoregulatory and anti-inflammatory effects, which have been reported to alleviate liver damage evoked by CCl4 or alcohol in previous reports. However, its chemopreventive effect against liver carcinogenesis is insufficient. The present study was aimed to investigate the possible role of MFP as a pro-apoptosis, and anti-inflammatory agent to possess its chemoprevention property. Hepatocarcinogenesis was induced by diethylnitrosamine/phenobarbital (DEN/PB) for 14 weeks. The DEN/PB-administered rats were co-treated with different doses of MFP (50 or 100 mg/kg body weight) by oral gavage for 14 weeks. Basic hepatic function indexes (AST, ALT, ALP, GGT, total bilirubin, and albumin), and hepatic tumor biomarkers (AFP, CEA, and CA19.9), together with histological assessment were performed. Besides, the hepatic apoptosis markers (Bcl-2, Bax, caspase3, and caspase9), inflammation markers (IL-1β, TNF-α, and NF-κB), and mutT homologue gene 1 (MTH1) were examined. Oral gavage of MFP inhibited the elevations of hepatic function indexes and hepatic tumor biomarkers and alleviated pathological changes in hepatic tissue. In addition, the hepatic apoptosis markers, inflammation markers, and the mRNA level of MTH1 were abnormal in DEN/PB group, which were reversed by MFP treatment. In conclusion, MFP is an effective agent that provides chemoprevention against DEN/PB-evoked hepatocarcinogenesis via inhibition of inflammation and induction of apoptosis.

to explore the value of transcatheter arterial chemoembolization (TACE) combined with targeted nanoparticle delivery system for sorafenib (SFB) to treat hepatocellular carcinoma (HCC) with microvascular invasion. 42 HCC patients with microvascular invasion after liver cancer surgery were selected from our hospital from December 2020 and February 2021. Patients were divided into experimental group and control group based on their willingness. Patients in experimental group (18 cases) were treated with combination therapy of TACE and Ab-SFB-NP system; while patients in control group (24 cases) took TACE and non-nano drug delivery system. There was no obvious difference in liver function and blood test results between two groups of patients before treatment and one month after treatment (P > 0.05). Three months after treatment, differences of alanine aminotransferase (ALT) were statistically significant (P 0.05). The disease control rate (DCR) of patients in experimental group was higher slightly (P > 0.05). The incidence of adverse reactions of patients in experimental group was lower than the control group and the differences were statistically significant (P

INTRODUCTION: Acute kidney injury is associated with high mortality in the perioperative period of liver transplantation. The aim of this position paper was to provide an up-to-date overview with special emphases on diagnosis, risk factors, and treatment. EVIDENCEACQUISITION: The Liver Intensive Care Group of Europe nominated a panel of recognized international experts who reviewed the available literature published from 1990 to January 2016 and produced clinical recommendations. The level of evidence and strength of recommendation were judged according to the Grading of Recommendations Assessment Development and Evaluation system. EVIDENCESYNTHESIS: Diagnosis of AKIshould be based on the KDIGOcriteria. The preoperative risk factors are more related to the patients predisposing factors and post-operative risk factors tend to be difficult to control. Therefore, focusing on intra-operative risk factors it would be important to maintain an adequate hemodynamics and to keep inferior vena cava clamping as short as possible. Biomarkers to identify AKIat an early stage are available; however, there is a lack of robust data that indicates their true beneficial effect. Intraoperative renal replacement therapy may be beneficial in some selective cases whereas its postoperative timing is still under debate. CONCLUSIONS: Perioperative liver transplant risk factors for acute kidney injury are difficult to control. Therefore, the focus should be on intra-operative hemodynamics and nephrotoxic drugs avoidance. Prospective randomized trials are needed to show the beneficial effect of early replacement therapy. In this context, the new biomarkers would be helpful in identifying kidney injury earlier OA embargo

The study reported in this paper tested the hypothesis that the developmental and reproductive health of mosquitofish (Gambusia affinis) exposed to pulp and paper effluent in the Dengcun River would differ from that of mosquitofish living in a reference site. We also studied whether morphological characteristics such as the anal fin and hemal spines of mosquitofish could serve as indicators for evaluating the androgenic effect and mosquitofish population security in the Dengcun River. Male and female mosquitofish were captured at three sites contaminated by pulp and paper effluent in the Dengcun River in Sihui, South China, and at a nearby uncontaminated reference site. Samples were collected from the sampling sites on the same day in August 2009. We compared the populations by total length, wet body and liver mass, gonad mass, and population composition. We also compared the populations according to number of anal fin segments, oocyte and embryo count, anal fin and hemal spine morphology among females, and by sperm count and viability among males, and observed the gonadal and liver histology of both males and females. Female mosquitofish exposed to pulp and paper effluent in the Dengcun River were generally smaller in length and mass, had a greater number of anal fin segments and more embryos, but had significantly fewer oocytes in comparison with those living at the reference site. The higher number of anal fin ray 3 segments and the increased ray 4:6 length ratio observed among fish taken from the Dengcun River sites indicated that they might be subject to the androgenic effect. Furthermore, the significantly different hemal spine morphology of the effluent-affected females also indicated the pulp and paper mills effluents in Dengcun River might contain androgenic substance(s). Male mosquitofish at the sites exposed to effluent had a higher number of anal fin segments and greater testis mass in comparison with those living at the reference site. No evidence of intersex was found in either males or females, although histopathological tests on females revealed histologic abnormalities in the liver and gonads. It can be concluded that pulp and paper effluent contamination in the Dengcun River has affected a number of developmental parameters and reproductive characteristics in mosquitofish, with possible adverse effects on reproduction in this population.

This review summarizes our current understanding of nonalcoholic fatty liver disease (NAFLD), a multi-factorial systemic disease resulting from a complex interaction between a specific genetic background and multiple environmental/metabolic “hits”. The role of gut microbiota, lipotoxicity, inflammation and their molecular pathways is reviewed in-depth. We also discuss the epidemiology and natural history of NAFLD by pinpointing the remarkably high prevalence of NAFLD worldwide and its inherent systemic complications: hepatic (steatohepatitis, advanced fibrosis and cirrhosis), cardio-metabolic (cardiovascular disease, cardiomyopathy, arrhythmias and type 2 diabetes) and neoplastic (primary liver cancers and extra-hepatic cancers). Moreover, we critically report on the diagnostic role of non-invasive biomarkers, imaging techniques and liver biopsy, which remains the reference standard for diagnosing the disease, but cannot be proposed to all patients with suspected NAFLD. Finally, the management of NAFLD is also reviewed, by highlighting the lifestyle changes and the pharmacological options, with a focus on the innovative drugs. We conclude that the results of ongoing studies are eagerly expected to lead to introduce into the clinical arena new diagnostic and prognostic biomarkers, prevention and surveillance strategies as well as to new drugs for a tailored approach to the management of NAFLD in the individual patient.