Detection of Frozen-Thawed Duck Fatty Liver by MALDI-TOF Mass Spectrometry: A Chemometrics Study.

The marketing of poultry livers is only authorized as fresh, frozen, or deep-frozen. The higher consumer demand for these products for a short period of time may lead to the marketing of frozen-thawed poultry livers: this constitutes fraud. The aim of this study was to design a method for distinguishing frozen-thawed livers from fresh livers. For this, the spectral fingerprint of liver proteins was acquired using Matrix-Assisted Laser Dissociation Ionization-Time-Of-Flight mass spectrometry. The spectra were analyzed using the chemometrics approach. First, principal component analysis studied the expected variability of commercial conditions before and after freezing-thawing. Then, the discriminant power of spectral fingerprint of liver proteins was assessed using supervised model generation. The combined approach of mass spectrometry and chemometrics successfully described the evolution of protein profile during storage time, before and after freezing-thawing, and successfully discriminated the fresh and frozen-thawed livers. These results are promising in terms of fraud detection, providing an opportunity for implementation of a reference method for agencies to fight fraud.

The Chinese Society of Hepatology position statement on the redefinition of fatty liver disease.

Fatty liver disease associated with metabolic dysfunction is of increasing concern in mainland China, the world's most populous country. The incidence of fatty liver disease is highest in China, surpassing the incidence in European countries and the USA. An international consensus panel recently published an influential report recommending a novel definition of fatty liver disease associated with metabolic dysfunction. This recommendation includes a switch in name from non-alcoholic fatty liver disease (NAFLD) to metabolic (dysfunction)-associated fatty liver disease (MAFLD) and adoption of a set of positive criteria for disease diagnosis that are independent of alcohol intake or other liver diseases. Given the unique importance of this proposal, the Chinese Society of Hepatology (CSH) invited leading hepatologists and gastroenterologists representing their respective provinces and cities to reach consensus on alternative definitions for fatty liver disease from a national perspective. The CSH endorses the proposed change from NAFLD to MAFLD (supported by 95.45% of participants). We expect that the new definition will result in substantial improvements in health care for patients and advance disease awareness, public health policy, and political, scientific and funding outcomes for MAFLD in China.

US-FLI score - Is it possible to predict the steatosis grade with an ultrasonographic score?

OBJECTIVES: A recent ultrasonographic score (Ultrasonographic fatty liver indicator (US-FLI)) allows to grade steatosis severity on ultrasound (US).We aimed to evaluate the agreement of US-FLI with the controlled attenuation parameter (CAP) in patients with non-alcoholic fatty liver disease (NAFLD). METHODS: Initially, inter-observer agreement for the score was assessed between 3 physicians using a sample of 31 patients.Later, 96 patients with NAFLD were included and several anthropometric/clinical/analytical parameters were assessed and US and transient elastography was performed. RESULTS: Physicians showed an excellent absolute agreement regarding the total score, with an average Interclass Correlation Coefficient of 0.972(95% CI 0.949-0.986). Comparing US-FLI with CAP, considering the previously defined cut-off for steatosis >S1(268dB/m) and > S2(280dB/m), US-FLI had a good discriminative capacity for both grades, with areas under the curve (AUC) of 0.88(p < 0.001) and 0.90(p < 0.001), respectively.Also, US-FLI ≤ 3 points had a negative predictive value of 100% for steatosis >S2 and US-FLI ≥6 points had a positive predictive value (PPV) of 94.0% for steatosis >S2. When comparing the clinical score Fatty Liver Index (FLI) for the same CAP cut-offs, it showed a weak discriminative capacity for both grades, with AUC of 0.65(p = 0.030) and 0.66(p = 0.017). AUC for US-FLI and FLI were significantly different for both cut-offs (p < 0.001). CONCLUSION: US-FLI has an excellent reproducibility and a good discriminative capacity for the different steatosis grades.Scores ≤3points exclude significant steatosis and scores ≥6 points have a PPV of 94,0% for steatosis >S2.US-FLI was significantly superior to the clinical score FLI in the discrimination between steatosis grades.

Estudio descriptivo de la esteatosis hepática y la morbilidad asociada en atención primaria. (Estudio ESTEATOAP) / Descriptive study of hepatic steatosis and associated morbidity in Primary Care. (ESTEATOAP Study)

OBJETIVO: Estudiar la prevalencia registrada de esteatosis hepática en atención primaria, así como la proporción de pacientes con diagnóstico de EHGNA que incluye el hígado graso simple no alcohólico (HGNA) y de esteatohepatitis no alcohólica frente a la esteatosis por otras causas. Además, se estudió la proporción de las morbilidades cardiometabólicas asociadas al HGNA. MATERIAL Y MÉTODOS: Estudio observacional descriptivo. La población de estudio es la de todos los pacientes con diagnóstico registrado de esteatosis hepática en un Centro de Salud urbano que atiende a una población de 25.747 mayores de 14 años. Se calculó un tamaño muestral de 229 pacientes. Se describen las características demográficas y clínicas asociadas. RESULTADOS: La prevalencia de esteatosis fue del 2,17%. Y de EHGNA del 1,51%. La media de edad de 62,42. Mujeres 114 mujeres (50,2%) y 113 varones (49,8%). Ciento cuarenta y siete (64,8%) fueron EHGNA y 64 (28,2%) fueron esteatosis por otras causas. La proporción de pacientes con EHGNA y transaminasas elevadas fue del 24,13% y la proporción de pacientes con EHGNA y elevación de GGT fue el 18,6%. Se ha encontrado una proporción elevada de EHGNA con factores de riesgo cardiometabólico: 93,9% de sobrepeso y obesidad, 55,1% de diabetes, 54,4% de hipertensión, 32,9% de síndrome metabólico, 35,2% hipertrigliceridemia y HDL de riesgo 19,6%. Entre los factores de riesgo cardiometabólicos y la EHGNA se encontró relación significativa en la diabetes, la obesidad y el síndrome metabólico. DISCUSIÓN: La prevalencia fue de solo el 1,51%, quizás por la escasa importancia que se da a esta enfermedad. Hay una proporción alta de EHGNA con factores de riesgo cardiometabólicos y más en la población general. Si se consideran todas las causas de esteatosis hay una asociación significativa entre obesidad, diabetes mellitus y síndrome metabólico con la EHGNA. CONCLUSIONES: La prevalencia registrada de EHGNA es muy inferior a la de los estudios poblacionales, además se ha encontrado una alta presencia de los factores cardiometabólicos en estos pacientes

OBJECTIVE: To study the recorded prevalence of hepatic steatosis in Primary Care, as well as the proportion of patients diagnosed with fatty liver diseases (FLD) including simple non-alcoholic fatty liver disease (NAFLD) and non-alcoholic steatohepatitis (NASH) versus steatosis by other causes. In addition, the proportion of cardiometabolic morbidities associated with NAFLD liver was studied. MATERIAL AND METHODS: A descriptive observational study was carried out on a population that included all patients with a recorded diagnosis of hepatic steatosis in an urban health centre that serves a population of 25,747 over the age of 14. A sample size of 229 patients was calculated. The demographic and clinical characteristics associated with hepatic steatosis are described. RESULTS: The prevalence of steatosis was 2.17% and 1.51% for NAFLD. The mean age was 62.42 years. The study included 114 (50.2%) women and 113 (49.8%) males. NAFLD was found in 147 (64.8%), and 64 (28.2%) were steatosis due to other causes. The proportion of patients with NAFLD and high transaminases was 24.13%, and the proportion of patients with NAFLD and GGT elevation was 18.6%. A high proportion of NAFLD had been found with cardiometabolic risk factors: 93.9% overweight and obesity, 55.1% diabetes, 54.4% hypertension, 32.9% metabolic syndrome, 35.2% hypertriglyceridaemia, and HDL risk 19.6%. A significant association was found between cardiometabolic risk factors and NAFLD in diabetes, obesity, and metabolic syndrome. DISCUSSION: Prevalence was only 1.51%, perhaps because of the low importance given to this disease. There is a high proportion of NAFLD with cardiometabolic risk factors and more in the general population. If all the causes of steatosis are considered there is a significant association between obesity, diabetes mellitus, and metabolic syndrome with NAFLD. CONCLUSIONS: The recorded prevalence of NAFLD is much lower than that of population studies, and a high presence of cardiometabolic factors has been found in these patients

A new definition for metabolic dysfunction-associated fatty liver disease: An international expert consensus statement.

The exclusion of other chronic liver diseases including "excess" alcohol intake has until now been necessary to establish a diagnosis of metabolic dysfunction-associated fatty liver disease (MAFLD). However, given our current understanding of the pathogenesis of MAFLD and its rising prevalence, "positive criteria" to diagnose the disease are required. In this work, a panel of international experts from 22 countries propose a new definition for the diagnosis of MAFLD that is both comprehensive and simple, and is independent of other liver diseases. The criteria are based on evidence of hepatic steatosis, in addition to one of the following three criteria, namely overweight/obesity, presence of type 2 diabetes mellitus, or evidence of metabolic dysregulation. We propose that disease assessment and stratification of severity should extend beyond a simple dichotomous classification to steatohepatitis vs. non-steatohepatitis. The group also suggests a set of criteria to define MAFLD-associated cirrhosis and proposes a conceptual framework to consider other causes of fatty liver disease. Finally, we bring clarity to the distinction between diagnostic criteria and inclusion criteria for research studies and clinical trials. Reaching consensus on the criteria for MAFLD will help unify the terminology (e.g. for ICD-coding), enhance the legitimacy of clinical practice and clinical trials, improve clinical care and move the clinical and scientific field of liver research forward.

Prevalence of steatosis and fibrosis in young adults in the UK: a population-based study.

BACKGROUND: The estimated worldwide prevalence of non-alcoholic fatty liver disease (NAFLD) in adults is 25%; however, prevalence in young adults remains unclear. We aimed to identify the prevalence of steatosis and fibrosis in young adults in a sample of participants recruited through the Avon Longitudinal Study of Parents and Children (ALSPAC), based on transient elastography and controlled attenuation parameter (CAP) score. METHODS: In this population-based study, we invited active participants of the ALSPAC cohort to our Focus@24+ clinic at the University of Bristol (Bristol, UK) between June 5, 2015, and Oct 31, 2017, for assessment by transient elastography with FibroScan, to determine the prevalence of steatosis and fibrosis. FibroScan data were collected on histologically equivalent fibrosis stage (F0-F4) and steatosis grade (S0-S3); results with an IQR to median ratio of 30% or greater were excluded for median fibrosis results greater than 7·1 kPa, and CAP scores for steatosis were excluded if less than ten valid readings could be obtained. Results were collated with data on serology (including alanine aminotransferase, aspartate aminotransferase, and γ-glutamyl transferase) and exposures of interest: alcohol consumption (via the Alcohol Use Disorder Identification Test for Consumption [AUDIT-C] and the Diagnostic and Statistical Manual of Mental Disorders-5 criteria for alcohol use disorder), body-mass index (BMI), waist-to-height ratio, socioeconomic status (based on predefined ALSPAC markers), and sex. We used logistic regression models to calculate odds ratios (ORs) for the effect of exposures of interest on risk of steatosis and fibrosis, after dichotomising the prevalences of fibrosis and steatosis and adjusting for covariates (excessive alcohol intake [hazardous drinking, AUDIT-C score ≥5; or harmful drinking, evidence of alcohol use disorder], social class, smoking, and BMI). FINDINGS: 10 018 active ALSPAC participants were invited to our Focus@24+ clinic, and 4021 attended (1507 men and 2514 women), with a mean age of 24·0 years (IQR 23·0-25·0). 3768 CAP scores were eligible for analysis. 780 (20·7% [95% CI 19·4-22·0]) participants had suspected steatosis (S1-S3; ≥248 dB/m), with 377 (10·0%) presenting with S3 (severe) steatosis (≥280 dB/m). A BMI in the overweight or obese range was positively associated with steatosis when adjusted for excessive alcohol consumption, social class, and smoking (overweight BMI: OR 5·17 [95% CI 4·11-6·50], p<0·0001; obese BMI: 27·27 [20·54-36·19], p<0·0001). 3600 participants had valid transient elastography results for fibrosis analysis. 96 participants (2·7% [95% CI 2·2-3·2]) had transient elastography values equivalent to suspected fibrosis (F2-F4; ≥7·9 kPa), nine of whom had values equivalent to F4 fibrosis (≥11·7 kPa). Individuals with alcohol use disorder and steatosis had an increased risk of fibrosis when adjusted for smoking and social class (4·02 [1·24-13·02]; p=0·02). INTERPRETATION: One in five young people had steatosis and one in 40 had fibrosis around the age of 24 years. The risk of fibrosis appears to be greatest in young adults who have harmful drinking patterns and steatosis. A holistic approach to the UK obesity epidemic and excessive drinking patterns is required to prevent an increasing health-care burden of adults with advanced liver disease in later life. FUNDING: Medical Research Council UK, Alcohol Change UK, David Telling Charitable Trust.

Hounsfield unit values of liver pathologies in unenhanced post-mortem computed tomography.

BACKGROUND: The goal of this study was to evaluate if unenhanced PMCT HU values of liver pathologies differ from post-mortem HU values of non-pathologic liver tissue. METHODS: Liver HU values were measured in five liver segments in PMCT unenhanced datasets of 214 forensic cases (124 male, 90 female, mean age 54.3 years). Liver HU values were compared with corresponding histologic liver findings. HU values of non-pathologic livers were compared to HU values of liver pathologies. RESULTS: A total of 64 non-pathologic livers (mean HU 58.32, SD 8.91) were assessed. Histologic diagnosed liver pathologies were as follows: steatosis (n = 121 (grade I n = 61, grade II n = 37, grade III n = 23)), fibrosis (n = 10), and cirrhosis (n = 19). HU values of the livers exhibiting severe steatosis (mean HU 32.44, SD 13.76), fibrosis (mean HU 44.7, SD 16.31), and cirrhosis (mean HU 50.59, SD 9.42) significantly differed to HU values of non-pathologic livers at ANOVA testing. CONCLUSION: PMCT unenhanced liver HU value measurements may be used as an additional method to detect unspecific liver-pathology. Values below 30 HU may specifically indicate severe steatosis.

Staging of Fatty Liver Diseases Based on Hierarchical Classification and Feature Fusion for Back-Scan-Converted Ultrasound Images.

Fatty liver disease is progressive and may not cause any symptoms at early stages. This disease is potentially fatal and can cause liver cancer in severe stages. Therefore, diagnosing and staging fatty liver disease in early stages is necessary. In this paper, a novel method is presented to classify normal and fatty liver, as well as discriminate three stages of fatty liver in ultrasound images. This study is performed with 129 subjects including 28 normal, 47 steatosis, 42 fibrosis, and 12 cirrhosis images. The proposed approach uses back-scan conversion of ultrasound sector images and is based on a hierarchical classification. The proposed algorithm is performed in two parts. The first part selects the optimum regions of interest from the focal zone of the back-scan-converted ultrasound images. In the second part, discrimination between normal and fatty liver is performed and then steatosis, fibrosis, and cirrhosis are classified in a hierarchical basis. The wavelet packet transform and gray-level co-occurrence matrix are used to obtain a number of statistical features. A support vector machine classifier is used to discriminate between normal and fatty liver, and stage fatty cases. The results of the proposed scheme clearly illustrate the efficiency of this system with overall accuracy of 94.91% and also specificity of more than 90%.

Molecular classification of fatty liver by high-throughput profiling of protein post-translational modifications.

We describe an alternative approach to classifying fatty liver by profiling protein post-translational modifications (PTMs) with high-throughput capillary isoelectric focusing (cIEF) immunoassays. Four strains of mice were studied, with fatty livers induced by different causes, such as ageing, genetic mutation, acute drug usage, and high-fat diet. Nutrient-sensitive PTMs of a panel of 12 liver metabolic and signalling proteins were simultaneously evaluated with cIEF immunoassays, using nanograms of total cellular protein per assay. Changes to liver protein acetylation, phosphorylation, and O-N-acetylglucosamine glycosylation were quantified and compared between normal and diseased states. Fatty liver tissues could be distinguished from one another by distinctive protein PTM profiles. Fatty liver is currently classified by morphological assessment of lipid droplets, without identifying the underlying molecular causes. In contrast, high-throughput profiling of protein PTMs has the potential to provide molecular classification of fatty liver.

La elastografía mediante técnica Acoustic radiation force impulse es eficaz en la detección de fibrosis hepática en el niño / Acoustic radiation force impulse imaging elastography is efficacious in detecting hepatic fibrosis in children

Objetivo. Evaluar el rendimiento diagnóstico de ARFI para detectar fibrosis hepática significativa en la edad pediátrica. Material y métodos. El estudio fue aprobado por el comité de ética hospitalario, con el consentimiento informado de los pacientes o sus representantes. Estudiamos 96 niños (50 varones, 46 hembras; edad media 8 años); 16 voluntarios sin enfermedad hepática conocida y 80 con patologías que pueden evolucionar a fibrosis y cirrosis hepática. La muestra final incluyó 31 pacientes con biopsia y 16 controles sanos. En todos los casos se realizó ecografía abdominal incluyendo Doppler y elastografía con ARFI. El valor ARFI expresado como velocidad (m/s) de propagación de las ondas transversales a través del tejido se calculó promediando 16 medidas en ambos lóbulos hepáticos. Comparamos las medias con el test de ANOVA de un factor. Los tests t de Student y chi cuadrado se usaron para datos categóricos. La significación estadística se estableció para una p < 0,05. Resultados. La velocidad en niños con fibrosis ≥ F2 fue significativamente más alta (1,80 ± 0,45 m/s) que en controles y pacientes con F0-F1 (1,38 ± 0,22 m/s) (p < 0,001). La esteatosis no se relacionó con la velocidad. La actividad necroinflamatoria se relacionó muy significativamente con la velocidad (p < 0,01). Fibrosis y actividad necroinflamatoria se relacionaron muy significativamente (p < 0,0001). Conclusión. La velocidad de propagación de las ondas ARFI se relacionó significativamente en los niños con el grado de fibrosis hepática (AU)

Objective. To evaluate the diagnostic performance of acoustic radiation force impulse imaging (ARFI) in detecting significant hepatic fibrosis in children. Material and methods. Our hospital's ethics committee approved the study and all patients or their representatives provided informed written consent. We included 96 children (50 boys, 46 girls; mean age, 8 y). We also studied 16 volunteers without liver disease as controls and 80 patients with diseases that can lead to fibrosis and cirrhosis of the liver. The final sample included 31 patients with biopsies and the 16 controls. All patients underwent abdominal ultrasonography including Doppler imaging and elastography with ARFI. The ARFI value, expressed as velocity (m/s) of shear wave propagation through the tissue, was calculated by averaging 16 measurements in both liver lobes. We used one-way analysis of variance to compare means between groups; we set statistical significance at P<.05. We used Student's t-tests and chi-square tests for categorical data. Results. The ARFI value in children with fibrosis ≥ F2 was higher (1.80 ± 0.45 m/s) than in controls and higher than in patients with F0-F1 (1.38 ± 0.22 m/s). The difference was significant (P<.001) for detecting F ≥ 2. Steatosis was not related with the ARFI value (Student's t-test, P>.84). Necroinflammatory activity was strongly associated with the ARFI value (Student's t-test, P<.01). Fibrosis and necroinflammatory activity were strongly associated with each other (chi-square test, P<.0001). Conclusion. The speed of shear wave propagation is significantly associated with the degree of hepatic fibrosis in children (AU)

Hígado graso no alcohólico en pacientes con sobrepeso y obesidad infantojuvenil / Non-alcoholic fatty liver in children and adolescents with excess weight and obesity

Fundamentos y objetivo: La esteatosis hepática, también denominada hígado graso no alcohólico (HGNA), es la enfermedad hepática más frecuente en niños obesos y está sufriendo un incremento importante en su incidencia debido al aumento de la obesidad infantil. Los objetivos de nuestro trabajo son: estimar la prevalencia de HGNA en niños con sobrepeso y obesidad utilizando el valor de la glutamate pyruvate transaminase (GPT, «transaminasa glutámico-pirúvica») y la ecografía abdominal, así como correlacionar la presencia de HGNA con distintos parámetros antropométricos y bioquímicos. Pacientes y método: Estudio transversal de prevalencia que incluye pacientes con sobrepeso y obesidad, con edades entre 5 y 15 años, reclutados entre 2004-2012. Las variables independientes incluidas fueron: edad, sexo, peso, talla, índice de masa corporal, perímetro de cintura (PC), índice cintura-talla (ICT), insulinemia,Homeostasis model assessment-insulin resistance (HOMA-R), colesterol total, triglicéridos (TG), high density lipoproteins (HDL, «lipoproteínas de alta densidad»), low density lipoproteins (LDL, «lipoproteínas de baja densidad»), glutamic-oxaloacetic transaminase (GOT, «transaminasa glutámico-oxalacética»), GPT y gamma glutamil transpeptidasa (GGT). Resultados: Se seleccionaron 126 pacientes, con una edad media (DE) de 11,94 (3,12) años. El 19,66% presentó elevación patológica de GPT. Treinta y ocho pacientes (30,15%) presentaron esteatosis hepática utilizando la ecografía abdominal. Los valores de insulinemia, HOMA-R y LDL fueron significativamente mayores en pacientes con alteración de GPT (p = 0,015, p = 0,008 y p = 0,002, respectivamente). Los pacientes con HGNA observado en ecografía también mostraron mayores valores de insulinemia, PC, ICT, colesterol total, TG, LDL, GOT, GPT y GGT que los pacientes con ecografía normal, alcanzándose la significación estadística en valor de insulinemia, HOMA, LDL y GPT. Conclusiones: La esteatosis hepática es un trastorno relativamente frecuente en niños y jóvenes con obesidad. Dos de cada 10 niños –utilizando GPT– y 3 de cada 10 –utilizando la ecografía abdominal– la presentan, y el marcador bioquímico que mejor la define es una elevación de la GPT. Es imprescindible y necesaria una modificación en el estilo de vida que incluya la pérdida de peso como medida principal para evitar complicaciones en la vida adulta (AU)

Basis and objective: Hepatic steatosis, also known as non-alcoholic fatty liver (NAFL), is the most frequent liver disease in obese children. Due to an increase in infantile obesity, it is experiencing a significant increment in incidence. Our objetives are: Estimate the prevalence of NAFL in children with excess weight and obesity using the glutamate pyruvate transaminase (GPT) value as a biochemical test and an abdominal ultrasound, and correlate the presence of hepatic steatosis with various anthropometric and biochemical parameters. Patients and method: Cross-sectional prevalence study which includes children with excess weight and obesity between the ages of 5 and 15 years, between the years 2004-2012. The independent variables included were: age, sex, weight, size, body mass index (BMI), waist circumference (WC), waist size index (WSI), insulinemia, Homeostasis model assessment-insulin resistance (HOMA-R), total cholesterol, triglycerides (TG), high density lipoproteins (HDL), low density lipoproteins (LDL), glutamic-oxaloacetic transaminase (GOT), GPT and gamma-glutamyl transpeptidase (GGT). Results: One hundred and twenty-six patients, with an average age of 11.94 (3.12) years were recruited. A percentage of 19.66 of the patients presented elevated GPT pathology. Of the 126 abdominal ultrasounds performed, 38 patients presented hepatic steatosis (30.15%). The levels of insulinemia, HOMA-R and LDL were significantly higher in patients with altered GPT, compared to those with normal GPT values (P = .015, P = .008 and P = .002, respectively). The patients with an objective HGNA in ultrasound, also showed greater levels of insulinemia, WC, WSI, total cholesterol, TG, LDL, GLT, GPT and GGT than the patients with normal ultrasounds, thereby achieving statistical significance in insulinemia, HOMA-R, LDL and GPT values. Conclusions: NAFL is a relatively frequent disorder in obese children and adolescents. In our study, 2 of 10 children –using GPT– and 3 of every 10 –using abdominal ultrasound– present the same. The biochemical marker which best defines it is an elevation in GPT. A modification in lifestyle which includes weight loss as a principal means of avoiding complications in adult life, is essential and necessary (AU)

Utility and appropriateness of the fatty liver inhibition of progression (FLIP) algorithm and steatosis, activity, and fibrosis (SAF) score in the evaluation of biopsies of nonalcoholic fatty liver disease.

UNLABELLED: Biopsy is still the gold standard for the diagnosis of nonalcoholic steatohepatitis but the definition may vary among pathologists, a drawback especially in evaluation of biopsies for clinical trials. We previously developed a scoring system (steatosis, activity, fibrosis [SAF]) allowing the use of an algorithm (fatty liver inhibition of progression [FLIP]) for the classification of liver injury in morbid obesity. The aim of this study was to determine whether the use of the SAF score and FLIP algorithm can decrease interobserver variations among pathologists. In a first session, pathologists categorized 40 liver biopsies of patients with nonalcoholic fatty liver disease (NAFLD) according to their own experience. In a second reading session, each pathologist reclassified the same slides by using the FLIP algorithm and SAF score, blinded to their first evaluation. The experiment was repeated with two different groups of pathologists at varying levels of training in liver pathology. The percentage of biopsy interpretation concordant with reference evaluation increased from 77% to 97% in Group 1 and from 42% to 75% in Group 2 after the use of the SAF score and FLIP algorithm. The strength of concordance in classification increased in Group 1 from moderate (κ = 0.54) to substantial (κ = 0.66) and from fair (κ = 0.35) to substantial (κ = 0.61) in Group 2 with application of the algorithm. With regard to the SAF score, concordance was substantial in Group 1 for steatosis (κ = 0.61), activity (κ = 0.75), and almost perfect for fibrosis (κ = 0.83 after pooling 1a, 1b, and 1c together into a single score F1). Similar trends were observed in Group 2 (κ = 0.54 for S, κ = 0.68 for A, and κ = 0.72 for F). CONCLUSION: The FLIP algorithm based on the SAF score should decrease interobserver variations among pathologists and are likely to be implemented in pathology practice.

Automatic classification of white regions in liver biopsies by supervised machine learning.

Automated assessment of histological features of non-alcoholic fatty liver disease (NAFLD) may reduce human variability and provide continuous rather than semiquantitative measurement of these features. As part of a larger effort, we perform automatic classification of steatosis, the cardinal feature of NAFLD, and other regions that manifest as white in images of hematoxylin and eosin-stained liver biopsy sections. These regions include macrosteatosis, central veins, portal veins, portal arteries, sinusoids and bile ducts. Digital images of hematoxylin and eosin-stained slides of 47 liver biopsies from patients with normal liver histology (n = 20) and NAFLD (n = 27) were obtained at 20× magnification. The images were analyzed using supervised machine learning classifiers created from annotations provided by two expert pathologists. The classification algorithm performs with 89% overall accuracy. It identified macrosteatosis, bile ducts, portal veins and sinusoids with high precision and recall (≥ 82%). Identification of central veins and portal arteries was less robust but still good. The accuracy of the classifier in identifying macrosteatosis is the best reported. The accurate automated identification of macrosteatosis achieved with this algorithm has useful clinical and research-related applications. The accurate detection of liver microscopic anatomical landmarks may facilitate important subsequent tasks, such as localization of other histological lesions according to liver microscopic anatomy.

Retrospective study of hepatocellular adenomas based on the phenotypic classification system: A report from China.

A molecular and pathological classification system for hepatocellular adenomas (HCAs) was recently introduced in Europe, resulting in four major identified subgroups. Asian countries have a considerably lower incidence of HCA as well as a different etiology. We aimed to characterize HCAs in a Chinese population based on this new classification system. A series of 30 patients with HCA were analyzed based on the phenotypic classification system using immunohistochemical analysis. Investigated antigens included liver-fatty acid binding protein (L-FABP), glutamine synthetase (GS), ß-catenin, serum amyloid A (SAA), and C-reactive protein (CRP). Of the 30 cases (20 female) included in this study, only one had a history of oral contraceptive use. We identified 9 (30%) hepatocyte nuclear factor (HNF)-1α-inactivated HCAs, 3 (10%) ß-catenin-activated HCAs, 11 (36.7%) inflammatory HCAs, and 7 (23.3%) unclassified HCAs. In the inflammatory HCA group, 2 cases demonstrated concurrent ß-catenin-activation. Homogeneous steatosis (6/9) and microadenomas (2/9) were more frequently observed in HNF1α-inactivated HCAs. A body mass index (BMI) of greater than 25 (5/11), alcohol use (4/11), and steatosis in background liver (3/11) were more frequent in inflammatory HCAs. ß-catenin-activated HCAs were larger than those of other subgroups. Despite obvious differences in etiology and gender proportion compared with Western countries, the clinical and pathological characteristics of HCA subgroups in China are similar to those in Europe. The phenotypic classification system could be reliably applied to Chinese patients as a meaningful tool for HCA management.

[Chronic liver diseases--new therapy. Are biopsies necessary?]. / Chronische Lebererkrankungen--neue Therapie. Benötigen wir die Biopsie?

Chronic liver disease can often reliably be assessed only by examination of biopsy material. In this article the possible indications for liver biopsy in viral hepatitis B and C, autoimmune liver disease, steatohepatitis and hereditary metabolic diseases are described. A biopsy may be useful in cases with unclear clinical or serological situations or with questionable chronicity and comorbidities. The assessment of biopsy material should be based on guideline-based classification systems. The value of biopsy diagnosis benefits from a close interdisciplinary clinical pathological cooperation.

Noninvasive predictors of nonalcoholic steatohepatitis in Korean patients with histologically proven nonalcoholic fatty liver disease.

BACKGROUND/AIMS: The aims of this study were (1) to identify the useful clinical parameters of noninvasive approach for distinguishing nonalcoholic steatohepatitis (NASH) from nonalcoholic fatty liver disease (NAFLD), and (2) to determine whether the levels of the identified parameters are correlated with the severity of liver injury in patients with NASH. METHODS: One hundred and eight consecutive patients with biopsy-proven NAFLD (age, 39.8±13.5 years, mean±SD; males, 67.6%) were prospectively enrolled from 10 participating centers across Korea. RESULTS: According to the original criteria for NAFLD subtypes, 67 patients (62.0%) had NASH (defined as steatosis with hepatocellular ballooning and/or Mallory-Denk bodies or fibrosis ≥2). Among those with NAFLD subtype 3 or 4, none had an NAFLD histologic activity score (NAS) below 3 points, 40.3% had a score of 3 or 4 points, and 59.7% had a score >4 points. Fragmented cytokeratin-18 (CK-18) levels were positively correlated with NAS (r=0.401), as well as NAS components such as lobular inflammation (r=0.387) and ballooning (r=0.231). Fragmented CK-18 was also correlated with aspartate aminotransferase (r=0.609), alanine aminotransferase (r=0.588), serum ferritin (r=0.432), and the fibrosis stage (r=0.314). A fragmented CK-18 cutoff level of 235.5 U/L yielded sensitivity, specificity, and positive and negative predictive values of 69.0%, 64.9%, 75.5% (95% CI 62.4-85.1), and 57.1% (95% CI 42.2-70.9), respectively, for the diagnosis of NASH. CONCLUSIONS: Serum fragmented CK-18 levels can be used to distinguish between NASH and NAFL. Further evaluation is required to determine whether the combined measurement of serum CK-18 and ferritin levels improves the diagnostic performance of this distinction.