Human Liver Specimens Research Articles

Morphological Variations and Measurements of the Caudate Lobe of the Human Liver: A Cadaveric Study

Abstract Background: The caudate lobe is an anatomic segment of the human liver and has a great clinical importance in hepatic diseases. Aim of Study: The aim of this study was to evaluate the morphological variations and anatomical measurements of the caudate lobe of the human liver that could be important in interpretation of its pathological conditions. Material and Methods: Measurements and morphological analysis of the caudate lobe of the human liver were determined in 56 formalin fixed human liver specimens in the Anatomy and Embryology Department, Faculty of Medicine, Zagazig University, Egypt. Results: The common shape of the caudate lobe was rectangular. The mean length of the caudate lobe was 57.45±4.74mm while the mean of its transverse diameter was 27.49±2.82mm. The mean of transverse diameter of the right lobe of the liver was 90.58±7.76mm and also, the mean of caudate lobe-right lobe (CL/RL) ratio was 0.30±0.03. Conclusion: Variable shapes of the caudate lobe of the liver were present. The ratio of the transverse diameter of caudate lobe to that of the right lobe of the liver ranged between 0.22-0.38. The collected data in this study may be valuable to aid the surgeons in proper planning of hepatic surgery and to avoid the misdiagnosis of hepatic diseases.

Grating-based phase-contrast CT (PCCT): histopathological correlation of human liver cirrhosis and hepatocellular carcinoma specimen

AimsTo correlate signal intensities in grating-based phase-contrast CT (PCCT) images obtained at a synchrotron light source and a conventional X-ray source with tissue components in human liver cirrhosis and hepatocellular...

Multispectral tissue mapping - a novel concept for the optical evaluation of liver malignancy

Introduction: Malignancy alters the optical absorption behaviour of liver tissue. This process can be evaluated using multi spectral analysis, a method that has been proposed as an intraoperative diagnostic tool. Existing systems either require invasive probe insertion or have limited spectral sensitivity which reduces their diagnostic ability. This work describes the development and evaluation of a novel, imaging method called multispectral tissue mapping, which addresses these issues. Methods: The imaging system (spectral range 1100-1700nm) consists of a tuneable excitation light source and a near-infrared camera. Following ethics approval, absorption spectra from resected human liver specimen were acquired ex-vivo. Twenty repeat acquisitions of malignant and non-malignant tissues from the same patient were averaged to reduce variability and subsequently compared. Generalised linear mixed modelling was employed for statistical analysis of serial changes in pixel based absorption (pixel area≈0.5mm2). Subsequently absorption data was transformed into augmented colour images with the aim of identifying cancer tissue. Results: Multiple spectra from liver specimens with colorectal cancer liver metastasis (CRLM) (three patients, n=505.141 pixel values) and hepatocellular carcinoma (HCC) (two patients, n=674.477 pixel values) were examined. Malignant tissue had a heterogeneous appearance on multispectral imaging compared to non-malignant tissue. Whereas absorption was increased in CRLM (maxima 1410nm; OR 0.005, CI 0.004-0.007) it was reduced in HCC (minimum 1440nm; OR 5.4, CI 3.9-7.6). Data was randomised into two groups and subsequently a predictive algorithm was built and tested. On ROC analysis predictive accuracy was 0.6 (area under curve) for HCC and 0.7 for CRLM, respectively. Augmented colour processing empirically enhanced image contrast between malignant and non-malignant tissue. Conclusion: Multispectral tissue mapping enables identification of liver malignancy on a sub-millimetre level. Augmented colour representation may enhance image contrast between malignant and normal liver tissues. Studies on larger datasets are required to improve the accuracy of the predictive algorithm.

A critical role of autophagy in regulating the mesenchymal transition of ductular cells in liver cirrhosis

Our previous studies have shown that autophagy mediates the link between ductular reaction (DR) and liver cirrhosis. Whether the subsequent fibrogenic response is regulated by increased autophagy in DR remains unclear. Here, using both human liver specimens and a rat model of liver cirrhosis induced by 2-acetylaminofluorene (AAF) and carbon tetrachloride (CCL4), we explored the involvement of autophagy in regulating mesenchymal transition of ductular cells. Ductular cells from AAF/CCL4 livers exhibited increased autophagy compared to those of normal livers. These cells showed morphological and functional characteristics of mesenchymal cells. Blocking autophagy using bafilomycin A1 or siRNA targeting ATG7 reduced the expression of mesenchymal markers in these ductular cells from AAF/CCL4 livers, indicating a role for autophagy in regulating the mesenchymal phenotype of ductular cells. Furthermore, we show that the mesenchymal transition in DR requires the activation of transforming growth factor-β (TGF-β) signaling in an autophagy-dependent manner. Importantly, in cirrhotic human livers, ductular cells that are positive for LC3B also showed increased expression of TGF-β and fibroblast-specific protein-1. Our data suggest activation of autophagy in ductular cells, and also demonstrate that it is required for the mesenchymal transition during the DR, processes that are critically involved in the pathogenesis of cirrhosis.

In vitro and in vivo human metabolism of a synthetic cannabinoid EAM-2201 detected by LC–quadrupole-ion trap-MS/MS and high-resolution LC–Orbitrap-MS/MS

The aim of this study is to characterize the metabolites of EAM-2201 in human hepatocytes obtained in vitro and those in liver and urine specimens obtained in vivo from the autopsy of an EAM-2201 abuser. EAM-2201 was incubated with human hepatocytes for 3 h in a CO2 incubator and the metabolites of EAM-2201 were produced. The human liver specimen was homogenized and the metabolites were extracted. The urine specimen was hydrolyzed first with β-glucuronidase and the metabolites were extracted. The tentative detection and identification of those metabolites were performed using liquid chromatography (LC)–quadrupole-ion trap tandem mass spectrometry (MS/MS) and high-resolution LC–Orbitrap-MS/MS, respectively. Twelve metabolites of EAM-2201 could be characterized in hepatocytes, liver and urine. They were produced by defluorination, hydroxylation, carboxylation, dehydrogenation, N-dealkylation and/or glucuronidation of EAM-2201. The most detectable metabolite obtained from the in vitro hepatocyte incubation was that monohydroxylated at the 4-ethylnaphthalene moiety of EAM-2201 after dehydrogenation. However, only a trace amount of the same metabolite was detected in the liver tissue obtained postmortem from an EAM-2201 abuser. In the liver tissue, the highest metabolite was 5-carboxypentyl derivative of EAM-2201, which was monohydroxylated at the 4-ethylnaphthalene moiety, followed by monohydroxylpentyl and monohydroxy-4-ethylnaphthalenyl derivative of EAM-2201 without dehydrogenation. Twelve metabolites of a synthetic cannabinoid, EAM-2201, were tentatively identified in human hepatocytes in vitro, and in human liver and urine specimens in vivo for the first time. There was a distinct difference in metabolism profile between the in vitro and in vivo results.

Ultrastructural Features of Human Liver Specimens from Patients Who Died of Dengue Hemorrhagic Fever.

Recent advances in electron microscopy and tomography have revealed distinct virus-induced endoplasmic reticulum (ER) structures unique for dengue virus (DV) and other flaviviruses in cell culture models, including hepatocytes. These altered ultrastructures serve as sites for viral replication. In this study, we used transmission electron microscopy to investigate whether such structures were present in the liver of fatal dengue hemorrhagic fever (DHF) autopsy cases. In parallel, electron microscopic examination of suckling mouse brains experimentally infected with DV was performed as an in vivo model of acute DV infection. Typical features of ER changes containing abundance of replicative virions were observed in neurons and microglia of DV-infected suckling mouse brains (SMB). This indicated that the in vivo DV infection could induce similar viral replication structures as previously described in the in vitro DV-infected cell model. Nevertheless, liver tissues from autopsy of patients who died of DHF showed scant changes of ER membrane structures and rare particles of virions in hepatocytes, despite overwhelming evidence for the presence of viral antigens and RNA–indicating active virus replication. Instead hepatocytes contained an abundance of steatotic vesicles and structural damages. This lack of structural changes indicative of virus replication in human hepatocytes is discussed.

Cystic artery: An anatomic morphological study and its clinical significance

Background Cystic artery is one of the arteries which is of utmost surgical importance, as it originates from the vast range of blood vessels, associated to the coeliac trunk and superior mesenteric artery and its relation to the biliary ducts is variable. Cystic artery is not only difficult to visualize, but difficult to approach during the surgery. This study was undertaken to study the anatomy of arterial variations of cystic artery in the specimens of liver and gallbladder by exploring the extrahepatic duct system, and in addition the relations of cystic artery in the Calot's triangle were also studied and observed. Materials and Methods The study was conducted on 50 human liver specimens with intact gallbladder and extrahepatic duct system, collected from the Department of Anatomy over a period of 5 years. The specimens obtained were fixed in 10% formalin and were finely dissected. The specimens were observed for parameters like the origin of the cystic artery, its length and diameter, mode and level of termination, relation to the Calot's triangle, and the extrahepatic duct system. Results Origin of the cystic artery was normal in 92% of cases and variations were seen in approximately 8% cases. The most common origin of the cystic artery was from the right hepatic artery, which was in 92% of the cases. In the present study, in 97% cases the cystic artery terminated by dividing into the superficial and deep branches. In the rest, the artery continued as a superficial branch, the deep branch being replaced by the accessory cystic artery. In 64% cases, the cystic artery was seen within the Calot's triangle, and in 36% of cases, it was outside the Calot's triangle. In 67% cases, the cystic artery was medial to the cystic duct, in approximately 63% cases the cystic artery was lateral to the common hepatic duct, and in 30% of the cases the cystic artery passed anterior to the cystic duct. Incidence of accessory cystic arteries in the present study was approximately 4%.

TRUSS Exacerbates NAFLD Development by Promoting IκBα Degradation in Mice.

There is no effective treatment method for nonalcoholic fatty liver disease (NAFLD), the most common liver disease. The exact mechanism underlying the pathogenesis of NAFLD remains to be elucidated. Here, we report that tumor necrosis factor receptor-associated ubiquitous scaffolding and signaling protein (TRUSS) acts as a positive regulator of NAFLD and in a variety of metabolic disorders. TRUSS expression was increased in the human liver specimens with NAFLD or nonalcoholic steatohepatitis, and in the livers of high-fat diet (HFD)-induced and genetically obese mice. Conditional knockout of TRUSS in hepatocytes significantly ameliorated hepatic steatosis, insulin resistance, glucose intolerance, and inflammatory responses in mice after HFD challenge or in spontaneous obese mice with normal chow feeding. All of these HFD-induced pathological phenotypes were exacerbated in mice overexpressing TRUSS in hepatocytes. We show that TRUSS physically interacts with the inhibitor of nuclear factor κB α (IκBα) and promotes the ubiquitination and degradation of IκBα, which leads to aberrant activation of nuclear factor κB (NF-κB). Overexpressing IκBαS32A/S36A , a phosphorylation-resistant mutant of IκBα, in the hepatocyte-specific TRUSS overexpressing mice almost abolished HFD-induced NAFLD and metabolic disorders. Conclusion: Hepatocyte TRUSS promotes pathological stimuli-induced NAFLD and metabolic disorders, through activation of NF-κB by promoting ubiquitination and degradation of IκBα. Our findings may provide a strategy for the prevention and treatment of NAFLD by targeting TRUSS.

A genome-wide association study identifies a susceptibility locus for biliary atresia on 2p16.1 within the gene EFEMP1.

Biliary atresia (BA) is a rare pediatric cholangiopathy characterized by fibrosclerosing obliteration of the extrahepatic bile ducts, leading to cholestasis, fibrosis, cirrhosis, and eventual liver failure. The etiology of BA remains unknown, although environmental, inflammatory, infectious, and genetic risk factors have been proposed. We performed a genome-wide association study (GWAS) in a European-American cohort of 343 isolated BA patients and 1716 controls to identify genetic loci associated with BA. A second GWAS was performed in an independent European-American cohort of 156 patients with BA and other extrahepatic anomalies and 212 controls to confirm the identified candidate BA-associated SNPs. Meta-analysis revealed three genome-wide significant BA-associated SNPs on 2p16.1 (rs10865291, rs6761893, and rs727878; P < 5 ×10−8), located within the fifth intron of the EFEMP1 gene, which encodes a secreted extracellular protein implicated in extracellular matrix remodeling, cell proliferation, and organogenesis. RNA expression analysis showed an increase in EFEMP1 transcripts from human liver specimens isolated from patients with either BA or other cholestatic diseases when compared to normal control liver samples. Immunohistochemistry demonstrated that EFEMP1 is expressed in cholangiocytes and vascular smooth muscle cells in liver specimens from patients with BA and other cholestatic diseases, but it is absent from cholangiocytes in normal control liver samples. Efemp1 transcripts had higher expression in cholangiocytes and portal fibroblasts as compared with other cell types in normal rat liver. The identification of a novel BA-associated locus, and implication of EFEMP1 as a new BA candidate susceptibility gene, could provide new insights to understanding the mechanisms underlying this severe pediatric disorder.

Bone Marrow-Derived Macrophage Contributes to Fibrosing Steatohepatitis Through Activating Hepatic Stellate Cells

Background & Aims: The role of macrophages in fibrosing steatohepatitis is largely unclear. We characterized the origin and molecular mechanisms of macrophages and its targeted therapy in fibrosing steatohepatitis. Methods: Fibrosing steatohepatitis was established in Alms1 mutant (foz/foz) and C57BL/6 wildtype mice fed high-fat/high-cholesterol (HFHC) or methionine-and choline-deficient (MCD) diet. Bone marrow transplantation (BMT) was performed to track the macrophage origin in fibrosing steatohepatitis. Macrophages were depleted by liposomal clodronate. Primary macrophages were isolated from bone marrow for adoptive transfer into mice. Results: Macrophage infiltration is significantly induced in two mouse models of fibrosing steatohepatitis and human NASH-fibrosis patients. Bone marrow-derived macrophages (BMMs) contribute to the hepatic macrophage accumulation in experimental fibrosing steatohepatitis. Hepatic BMMs depletion by liposomal clodronate significantly attenuated fibrosing steatohepatitis. This effect was associated with reduced hepatic stellate cell (HSC) activation, collagen deposition and hepatic expression of key pro-fibrotic factors (TIMP1, TIMP2 and TGFβ1) and endoplasmic reticulum (ER) stress markers (GRP78, IRE1α and PDI). Conversely, adoptive transfer of BMMs significantly aggravated fibrosing steatohepatitis. Moreover, macrophage conditioned medium directly promoted the phenotypic transition of primary quiescent HSCs to activated HSCs; enhanced activation and proliferation (P<0.01) but decreased apoptosis (P<0.001) of HSC cell lines (LX-2 and HSC-T6). The effect of BMMs in promoting fibrosing steatohepatitis was mediated by inducing key pro-fibrosis factors and signaling pathways including cytokine/chemokine, TGF-β, and complement cascade by cDNA expression array. Complement 3a receptor (C3aR1) was a predominant effector of macrophage mediated fibrosing steatohepatitis. Knockout C3aR1 in mice blunted the development of fibrosing steatohepatitis. Conclusions: Macrophages recruited from bone marrow promote the progression of fibrosing steatohepatitis through directly activating HSCs. BMMs targeted therapy is a promising approach for fibrosing steatohepatitis. Funding: The project was supported by the RGC-GRF Hong Kong (14106415, 14111216, 14163817); Science and Technology Program Grant Shenzhen (JCYJ20170413161534162); Vice-Chancellor's Discretionary Fund CUHK; Shenzhen Virtual University Park Support Scheme to CUHK Shenzhen Research Institute; CUHK Direct grant. The human liver specimens were provided by Prof. Vincent WS Wong in the Prince of Wales Hospital, the Chinese University of Hong Kong and Prof. Kwan Man in the Queen Mary Hospital, University of Hong Kong, Hong Kong. Competing interests: All authors declare that they have no competing interests. Ethics Approval Statement: All subjects were given written informed consent prior to sample collection and the study protocol was approved by the Clinical Research Ethics Committee of the Chinese University of Hong Kong.

Expression of Multiple Sulfotransferase (SULT) 1C4 Transcript Variants in Developing Human Liver

The cytosolic sulfotransferases (SULTs) are conjugating enzymes that catalyze the transfer of a sulfonate group from the cofactor 3′‐phosphoadenosine‐5′‐phosphosulfate to a variety of endogenous and xenobiotic substrates. The SULT1C subfamily includes three human genes, SULT1C2, SULT1C3, and SULT1C4, and relatively little is known about their expression and regulation or the functions of the enzymes. Previous studies have indicated that the human SULT1Cs are preferentially expressed during early life and are capable of metabolizing endogenous molecules such as thyroid and estrogen hormones, suggesting that these enzymes could play physiological roles during development. Several transcript variants (TVs) of SULT1C4 are indexed in GenBank, including the full‐length mRNA containing seven exons (TV1, NM_006588), a variant mRNA lacking exons 3 and 4 (TV2, NM_001321770), two non‐coding RNA variants (TV3, NR_135776 and TV4, NR_135779), and a predicted transcript variant (TVX1, XM_017003807). Using 5′‐RACE, RT‐PCR, and DNA sequencing, we detected TV1 and TV2, as well as a variant lacking only exon 3 (E3DEL) in Caco‐2 colorectal adenocarcinoma cells, HepaRG hepatic cells, and human liver specimens. The aim of the current study was to evaluate expression of the SULT1C4 TVs more thoroughly by (1) characterizing expression of individual SULT1C4 TVs in human liver specimens from prenatal, infant, and adult donors and (2) determining whether the individual mRNA variants can be translated into proteins. Using RT‐qPCR assays designed to quantify TV1, TV2, or E3DEL separately, all three variants were found to be primarily expressed in prenatal liver samples. The TV2 transcript lacking two exons was more abundant than the full‐length TV1, while E3DEL levels were minimal. In agreement with the RT‐qPCR data, RNA‐seq analysis of prenatal and pediatric liver specimens showed that TV1 and TV2 were both more abundant during early developmental stages. The RNA‐seq data also demonstrated that a non‐coding transcript variant (ENST00000494122.1) consisting of 2 exons only was expressed in the liver specimens and was predominantly expressed in prenatal liver. TV1 and TV2, but not E3DEL, were detected by western blot when plasmids expressing DDK‐tagged variants were transfected into HEK‐293T cells. These results indicate that at least four transcript variants of SULT1C4 are expressed in human liver, primarily during prenatal life, although only one of these (TV1) would encode a functional SULT enzyme. The physiological significance of this transcript diversity is unclear and will require further investigation.Support or Funding InformationSupported by NIH grants R01 ES022606 and P30 ES020957This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Altered AKAP12 expression in portal fibroblasts and liver sinusoids mediates transition from hepatic fibrogenesis to fibrosis resolution

Liver fibrosis can be reversed by removing its causative injuries; however, the molecular mechanisms mediating the resolution of liver fibrogenesis are poorly understood. We investigate the role of a scaffold protein, A-Kinase Anchoring Protein 12 (AKAP12), during liver fibrosis onset, and resolution. Biliary fibrogenesis and fibrosis resolution was induced in wild-type (WT) or AKAP12-deficient C57BL/6 mice through different feeding regimens with 0.1% 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC)-containing chow. AKAP12 expression in portal fibroblasts (PFs) and liver sinusoidal endothelial cells (LSECs) gradually decreased as fibrosis progressed but was restored after cessation of the fibrotic challenge. Histological analysis of human liver specimens with varying degrees of fibrosis of different etiologies revealed that AKAP12 expression diminishes in hepatic fibrosis from its early stages onward. AKAP12 KO mice displayed reduced fibrosis resolution in a DDC-induced biliary fibrosis model, which was accompanied by impaired normalization of myofibroblasts and capillarized sinusoids. RNA sequencing of the liver transcriptome revealed that genes related to ECM accumulation and vascular remodeling were mostly elevated in AKAP12 KO samples. Gene ontology (GO) and bioinformatic pathway analyses identified that the differentially expressed genes were significantly enriched in GO categories and pathways, such as the adenosine 3′,5′-cyclic monophosphate (cAMP) pathway. Knockdown of the AKAP12 gene in cultured primary PFs revealed that AKAP12 inhibited PF activation in association with the adenosine 3′,5′-cyclic monophosphate (cAMP) pathway. Moreover, AKAP12 knockdown in LSECs led to enhanced angiogenesis, endothelin-1 expression and alterations in laminin composition. Collectively, this study demonstrates that AKAP12-mediated regulation of PFs and LSECs has a central role in resolving hepatic fibrosis.

Nrf2 induces lipocyte phenotype via a SOCS3-dependent negative feedback loop on JAK2/STAT3 signaling in hepatic stellate cells

Hepatic stellate cells (HSCs) are universally acknowledged to play a core role in the pathogenesis of hepatic fibrosis. HSCs when activated are characterized by dramatic loss of intracellular lipid droplets. Accumulative evidence has suggested that recovery of lipid droplets could suppress HSC activation. However, the underlying molecular mechanisms still remain largely unclear. In this study, we found that the expression and activity of nuclear factor (erythroid-derived 2) - like 2 (Nrf2) were decreased in activated HSCs and negatively correlated with hepatic fibrosis severity in human liver specimens. Nrf2 overexpression, in contrast to Nrf2 deficiency, induced the accumulation of lipid droplets via decreasing the expression of lipolytic gene peroxisome proliferator-activated receptor alpha (PPARα) and increasing the expression of genes involved in lipogenesis and retinoic acid responsiveness, including CCAAT/enhancer-binding protein alpha, PPARγ, retinoid X receptor alpha, and retinoic acid receptor beta. Consequently, HSCs regained its lipocyte phenotype and expressed reduced alpha-smooth muscle actin and collagen type I. Consistently, disruption of Janus kinase 2/signal transducer and activator of transcription 3 (JAK2/STAT3) signaling by AG490 or Stattic could also induce lipocyte phenotype. Noticeably, Nrf2 overexpression by a genetic approach disrupted JAK2/STAT3 signaling and increased the expression of suppressor of cytokine signaling 3 (SOCS3) but not other protein inhibitors of activated STATs. Gain- or loss-of function of SOCS3 revealed that Nrf2 inhibited JAK2/STAT3 signaling via inducing SOCS3 expression. In conclusion, Nrf2 activation induced lipocyte phenotype in HSCs via enhancing SOCS3-dependent feedback inhibition on JAK2/STAT3 cascade. Nrf2 could be a target molecule for antifibrotic strategy.

Methyleugenol DNA adducts in human liver are associated with SULT1A1 copy number variations and expression levels

Methyleugenol is a rodent hepatocarcinogen occurring in many herbs and spices as well as essential oils used for flavoring. Following metabolic activation by cytochromes P450 (CYPs) and sulfotransferases (SULTs), methyleugenol can form DNA adducts. Previously, we showed that DNA adduct formation by methyleugenol in mouse liver is dependent on SULT1A1 expression and that methyleugenol DNA adducts are abundant in human liver specimens. In humans, SULT1A1 activity is affected by genetic polymorphisms, including single-nucleotide polymorphisms (SNPs) and copy number variations (CNVs). Here we investigated the relationship between individual methyleugenol DNA adduct levels and SULT1A1 in human liver samples. Using isotope-dilution ultraperformance liquid chromatography coupled with tandem mass spectrometry, we quantified methyleugenol DNA adducts in 121 human surgical liver samples. Frequent CNVs, including deletions (f = 3.3%) and duplications (f = 36.4%) of SULT1A1, were identified using qPCR and TaqMan assays in the donors' genomic DNA. SULT1A1 mRNA and protein levels were quantified using microarray data and Western blot analysis, respectively. Methyleugenol DNA adducts were detected in all 121 liver samples studied. Their levels varied 122-fold between individuals and were significantly correlated to both mRNA and protein levels of SULT1A1 (r s = 0.43, and r s = 0.44, respectively). Univariate and multivariate statistical analysis identified significant associations of SULT1A1 CNVs with mRNA (p = 1.7 × 10-06) and protein (p = 4.4 × 10- 10) levels as well as methyleugenol DNA adduct levels (p = 0.003). These data establish the importance of SULT1A1 genotype for hepatic methyleugenol DNA adducts in humans, and they confirm a strong impact of SULT1A1 CNVs on SULT1A1 hepatic phenotype.

Changes in human hepatic metabolism in steatosis and cirrhosis.

AIMTo understand the underlying metabolic changes in human liver disease we have applied nuclear magnetic resonance (NMR) metabolomics analysis to human liver tissue.METHODSWe have carried out pilot study using 1H-NMR to derive metabolomic signatures from human liver from patients with steatosis, nonalcoholic steatohepatitis (NASH) or alcohol-related liver damage (ARLD) to identify species that can predict outcome and discriminate between alcohol and metabolic-induced liver injuries.RESULTSChanges in branched chain amino acid homeostasis, tricarboxylic acid cycle and purine biosynthesis intermediates along with betaine were associated with the development of cirrhosis in both ARLD and nonalcoholic fatty liver disease. Species such as propylene glycol and as yet unidentified moieties that allowed discrimination between NASH and ARLD samples were also detected using our approach.CONCLUSIONOur high throughput, non-destructive technique for multiple analyte quantification in human liver specimens has potential for identification of biomarkers with prognostic and diagnostic significance.

Morphological study of anomalies of lobes and Segments of hu man liver

Objective: To identify various morphological variations in surface of human liver. Materials and Methods: The study was conducted in 60 adult human liver specimens by dissection method. Results: out of 60 specimens studied 25cases (41%) showed variations. Presence of additional lobe in 3 cases (5%); accessory fissures in 8 cases (13%); short gall bladder in 3 cases (5%); elongated left lobe in 2 cases (3.3%);very small left lobe in 1 case (1.6%);absence of quadrate lobe in 2 cases (3.3%);very small quadrate lobe in 1 case (1.6%);absence of fissure for ligamentum teres 1 case (1.6%); incomplete fissure for ligamentum teres in 3 cases (5%); prominent caudate process in 1 case (1.6%).Conclusion: Thorough knowledge of these variations is a must for surgeons for planning any surgeries and to radiologist for correct interpretation of diagnosis and treatment occurring in that region.

Sex-determining region Y-box 9 acts downstream of NADPH oxidase to influence the effect of leptin on PPARγ1 expression in hepatic stellate cells

Leptin, an adipocyte-derived hormone, promotes liver fibrogenesis and inhibits the expression of peroxisome-proliferator activated receptor γ (PPARγ), a key transcription factor in inhibition of hepatic stellate cell (HSC) activation, in HSCs. This research aimed to further investigate the mechanisms underlying leptin regulation of PPARγ1 in HSCs in vivo and in vitro. Results demonstrated that sex-determining region Y-box 9 (Sox9) could bind to a site around −2275 within leptin response region of PPARγ1 promoter and inhibited PPARγ1 expression. Sox9 upregulated the expressions of α1(I)collagen and alpha-smooth muscle actin in HSCs. Leptin stimulated Sox9 expression and Sox9 binding to PPARγ1 promoter. The signaling pathways of NADPH oxidase, β-catenin, and delta-like homolog1 (DLK1) mediated leptin upregulation of Sox9 expression. Moreover, there existed crosstalk between NADPH oxidase pathway and β-catenin or DLK1 signaling pathway. Human liver specimens of cirrhosis were shown to be of a large number of the positive HSCs for p47phox (playing a central role in NADPH oxidase activity), 4-hydroxynonenal (a lipid peroxidation product), Sox9, and α-smooth muscle actin whereas PPARγ-positive HSCs were rarely detected. These results might deepen understanding of the molecular mechanisms for leptin inhibition of PPARγ1 expression in HSCs and for the liver fibrosis associated with leptin.

Human Ex-Vivo Liver Model for Acetaminophen-induced Liver Damage.

Reliable test systems to identify hepatotoxicity are essential to predict unexpected drug-related liver injury. Here we present a human ex-vivo liver model to investigate acetaminophen-induced liver injury. Human liver tissue was perfused over a 30 hour period with hourly sampling from the perfusate for measurement of general metabolism and clinical parameters. Liver function was assessed by clearance of indocyanine green (ICG) at 4, 20 and 28 hours. Six pieces of untreated human liver specimen maintained stable liver function over the entire perfusion period. Three liver sections incubated with low-dose acetaminophen revealed strong damage, with ICG half-lives significantly higher than in non-treated livers. In addition, the release of microRNA-122 was significantly higher in acetaminophen-treated than in non-treated livers. Thus, this model allows for investigation of hepatotoxicity in human liver tissue upon applying drug concentrations relevant in patients.

Abstract 3594: The TGF-β effector β2SP depletion abrogates DNA damage repair

Abstract Objective: Exposure to genotoxins, such as ethanol-derived acetaldehyde, leads to DNA damage, liver injury, and promotes the development of cancer. Alcohol-related genotoxicity, arising from DNA damage by metabolically generated reactive aldehydes, has recently been observed in models with genetic inactivation of members of the Fanconi anemia pathway. However, sensors for genotoxicity leading to aberrant DNA repair remain elusive. Transforming growth factor β (TGF-β) is a critical protein in the regulation of several cancer phenotypes and also functions as an extracellular sensor of ionizing radiation-induced cell damage. Yet, how the TGF-β pathway contributes to toxin-induced DNA damage repair remains unclear. We utilized the TGF-β/β2SP mutant mouse model to investigate the mechanisms in relation to β2SP-mediated TGF-β modulation of the Fanconi anemia pathway for DNA damage repair, alcohol sensitivity, and liver tumorigenesis. Methods: (1) β2SP mutant mice were treated with alcohol to determine their susceptibility to aldehyde-induced developmental abnormalities. (2) Genomic instability and sensitivity to DNA damaging agents in primary β2SP+/+, β2SP-/- MEFs were determined by clonogenic survival and metaphase chromosome aberrations analysis. (3) Defective S-phase specific DNA repair in β2SP-/- MEFs were determined by DNA replication restart assays. (4) ChIP assays were performed to determine the recruitment of β2SP/Smad3 at FancD2 promoter. (5) We investigated the clinical relevance of altered β2SP and FancD2 function using immunohistochemical analyses of 20 human liver specimens from alcoholic hepatitis (n = 5), alcoholic cirrhosis (n = 5), and alcohol-associated liver cancer (n = 5), as well as normal controls (n = 5). Results: (1) Sptbn1-deficient mice exhibit a phenotype similar to human fetal alcohol syndrome and are sensitive to ethanol exposure. (2) Sptbn1-deficient cells exhibit genomic instability and hypersensitivity to DNA damage (3) Sptbn1-deficiency delays DNA damage repair. (4) Furthermore, Sptbn1-deficient cells are defective in stalled DNA replication fork resolution and homologous recombination. (5) FancD2 ectopic expression rescues the DNA repair defect in Sptbn1 null cells. (6) β2SP and FancD2 are clinically correlated in alcoholic hepatitis and HCCs. Conclusions: Our model proposes that in response to liver toxins such as alcohol, the TGF-β/β2SP/Smad3 pathway prevents liver injury and cancer through its direct effects on DNA repair and genomic stability. Thus, characterizing the role of TGF-β in alcohol-induced injury could potentially enhance our mechanistic insight into the basis for therapeutics targeting toxin-induced DNA damage and tumorigenesis. Citation Format: Jian Chen, Vivek Shukla, Jiun-Sheng Chen, Raj K. Panditab, Yun Seong Jeong, Lior H Katz, Ji-Hyun Shin, YoungJin Gi, Lawrence N. Kwongc, Clayton R. Huntb, Patrizia Farci, Xiaoping Su, Jon White, Bibhuti Mishra, Asif Rashid, Milind Javle, Lei Li, Junjie Chen, John R, Stroehlein, Marta Davila, Rehan Akbani, Keigo Machidao, Hidekazu Tsukamoto, Tej K. Pandita, Lopa Mishra. The TGF-β effector β2SP depletion abrogates DNA damage repair. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3594.

Left Hemiliver Medial Sector-Portal Venous Anatomy

Background: The aim of this study was to classify the left hemiliver medial sector portal branches on the basis of their site of origin and area that supply. The leading motive for this investigation was to contribute to the modern liver practice, especially to the respective and transplantation ones. Methods: The examined material consisted of 27 portobiliary casts performed by injection-corrosive method using adult cadaveric human liver specimens. During observation under magnifying lens were determined the site and kind of origin of medial sector portal branches, numerical frequency of segmental portobiliary elements and their relation. Results: According to the supplied area two distinct segmentssuperior IVa and inferior IVb were distinguished. At the level of each segment portal branches appeared as main, additional and newly ramified. As to their origin at the level of segment IVa the mixed origin (separate and common) was prevalent versus separate or only common origin - 17/7/2 cases and portal branches were not observed in only one case in this segment. Also, at the level of segment IVb the mixed origin of portal branches was prevalent versus separate ones - 19/8 cases. The incidence of newly ramified branches to segment IVa was 22.22% and to segment IVb 37.037%. At the level of segment IVa there were additional or newly ramified branches on the superior surface of umbilical portion of the left portal vein branch (37.037%) and only additional branches for segment IVb on the superior, as well as inferior surface of this portion (7.41%). Conclusion: In the left hemiliver medial sector portal branches supply two distinct segments-superior SgIVa and inferior SgIVb. At the level of each segment multiple branches with varied number appear as main, additional and newly ramified. According to their origin the branches may be either separate for each segment or additionally to the branches with separate origin common branches for both segments and only with common origin branches to the SgIVa.