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Background: This study evaluated electrical conductivity in human liver tissue in the 3–1000 kHz frequency range to compare normal versus tumor tissues under in vivo versus ex vivo conditions. Methods: Previous informed consent was obtained from twenty patients undergoing liver resection in whom liver electrical conductivity was measured during surgery and after resection. Result: We found higher electrical conductivity values in tumor tissues than in normal tissue in both in vivo (0.41 ± 0.10 vs. 0.13 ± 0.06 S/m) and ex vivo (0.27 ± 0.09 vs. 0.12 ± 0.07 S/m) conditions (at 3 kHz). The electric properties also showed a promising potential for distinguishing between different tissue types including metastasis, cholangiocarcinoma (CCA), hepatocellular carcinoma (HCC), hepatic cirrhosis, and normal liver (both in vivo and ex vivo). At 3 kHz, in vivo electrical conductivity for cholangiocarcinoma, HCC, and metastasis were 0.35, 0.42 ± 0.13, and 0.41 ± 0.08 S/m, respectively, which differed significantly from each other (p < 0.05). Conclusions: These findings could potentially improve liver disease diagnostics through electrical conductivity measurements and treatment techniques involving electric fields. Future research should focus on expanding the sample size to refine the categorization and comparison processes across diverse human liver tissue types. [ABSTRACT FROM AUTHOR]

Neural regulation of hepatic metabolism has long been recognized. However, the detailed afferent and efferent innervation of the human liver has not been systematically characterized. This is largely due to the liver's high lipid and pigment contents, causing false-negative (light scattering and absorption) and false-positive (autofluorescence) results in in-depth fluorescence imaging. Here, to avoid the artifacts in three-dimensional (3-D) liver neurohistology, we embed the bleached human liver in the high-refractive-index polymer for tissue clearing and antifade 3-D/Airyscan super-resolution imaging. Importantly, using the paired substance P (SP, sensory marker) and PGP9.5 (pan-neuronal marker) labeling, we detect the sensory nerves in the portal space, featuring the SP+ varicosities in the PGP9.5+ nerve bundles/fibers, confirming the afferent liver innervation. Also, using the tyrosine hydroxylase (TH, sympathetic marker) labeling, we identify 1) condensed TH+ sympathetic nerves in the portal space, 2) extension of sympathetic nerves from the portal to the intralobular space, in which the TH+ nerve density is 2.6 ± 0.7-fold higher than that of the intralobular space in the human pancreas, and 3) the TH+ nerve fibers and varicosities contacting the ballooning cells, implicating potential sympathetic influence on hepatocytes with macrovesicular fatty change. Finally, using the vesicular acetylcholine transporter (VAChT, parasympathetic marker), PGP9.5, and CK19 (epithelial marker) labeling with panoramic-to-Airyscan super-resolution imaging, we detect and confirm the parasympathetic innervation of the septal bile duct. Overall, our labeling and 3-D/Airyscan imaging approach reveal the hepatic sensory (afferent) and sympathetic and parasympathetic (efferent) innervation, establishing a clinically related setting for high-resolution 3-D liver neurohistology. NEW & NOTEWORTHY We embed the human liver (vs. pancreas, positive control) in the high-refractive-index polymer for tissue clearing and antifade 3-D/Airyscan super-resolution neurohistology. The pancreas-liver comparison reveals: 1) sensory nerves in the hepatoportal space; 2) intralobular sympathetic innervation, including the nerve fibers and varicosities contacting the ballooning hepatocytes; and 3) parasympathetic innervation of the septal bile duct. Our results highlight the sensitivity and resolving power of 3-D/Airyscan super-resolution imaging in human liver neurohistology. [ABSTRACT FROM AUTHOR]

Objective：To discuss the effect of apolipoprotein C1 (APOC1) expression on the proliferation and apoptosis of the hepatocellular carcinoma cells, and to preliminarily clarify the related molecular mechanism. Methods：The expression level of APOC1 mRNA in hepatocellular carcinoma tissue and its relationship with the prognosis of the patient were analyzed by The Cancer Genome Atlas (TCGA) Database； real-time fluorescence quantitative PCR (RT-qPCR) method was used to detect the expression levels of APOC1 mRNA in different hepatocellular carcinoma cells； the human liver cancer HepG2 cells with low APOC1 expression were selected as the subjects. The HepG2 cells were transfected with pcDNA3. 1-APOC1 plasmid to over-express APOC1（APOC1 over-expression group), and the HepG2 cells transfected with empty vector pcDNA3. 1 were regarded as control group. MTS assay and 5-ethynyl2'-deoxyuridine (EdU) staining were used to detect the proliferative activities and proliferation rates of the cells in two groups； Transwell chamber assay was used to detect the numbers of migration cells in two groups；flow cytometry and TUNEL assay were used to detect the percentages of the cells at different cell cycles and apoptotic rates in two groups；Western blotting method was used to detect the expression levels of extracellular regulated protein kinase (ERK), phosphorylated ERK (p-ERK), protein kinase B (AKT), phosphorylated AKT (p-AKT), B-cell lymphoma-2 (Bcl-2), and cleaved cysteinyl aspartate specific proteinase-3 (cleaved caspase-3) proteins in the cells in two groups. Results：The TCGA Database results showed that the expression level of APOC1 mRNA in hepatocellular carcinoma tissue was lower than that in normal liver tissue (P<0. 05), and the patients with low expression of APOC1 mRNA had poor prognosis. The RT-qPCR results showed that the expression level of APOC1 mRNA in the HepG2 cells was the lowest, and the HepG2 cells were chosen for the subsequent research. Compared with control group, the proliferative activity and proliferation rate of the cells in APOC1 over-expression group were decreased (P<0. 05 or P<0. 01), the number of migration cells was decreased (P<0. 01), and the percentage of the cells at S phase and the apoptotic rate were significantly increased (P<0. 01) . Compared with control group, the expression levels of p-ERK, p-AKT, and Bcl-2 proteins in the cells in APOC1 over-expression group were significantly decreased (P<0. 05), and the expression level of cleaved caspase-3 protein was increased (P<0. 01) . Conclusion：High expression of APOC1 can inhibit the proliferation of the human liver cancer HepG2 cells and induce the apoptosis, and its mechanism may be related to inhibition of the expressions of p-ERK, p-AKT, Bcl-2 proteins and promotion of the expression of cleaved caspase-3 protein. [ABSTRACT FROM AUTHOR]

Background: This study evaluated electrical conductivity in human liver tissue in the 3–1000 kHz frequency range to compare normal versus tumor tissues under in vivo versus ex vivo conditions. Methods: Previous informed consent was obtained from twenty patients undergoing liver resection in whom liver electrical conductivity was measured during surgery and after resection. Result: We found higher electrical conductivity values in tumor tissues than in normal tissue in both in vivo (0.41 ± 0.10 vs. 0.13 ± 0.06 S/m) and ex vivo (0.27 ± 0.09 vs. 0.12 ± 0.07 S/m) conditions (at 3 kHz). The electric properties also showed a promising potential for distinguishing between different tissue types including metastasis, cholangiocarcinoma (CCA), hepatocellular carcinoma (HCC), hepatic cirrhosis, and normal liver (both in vivo and ex vivo). At 3 kHz, in vivo electrical conductivity for cholangiocarcinoma, HCC, and metastasis were 0.35, 0.42 ± 0.13, and 0.41 ± 0.08 S/m, respectively, which differed significantly from each other (p < 0.05). Conclusions: These findings could potentially improve liver disease diagnostics through electrical conductivity measurements and treatment techniques involving electric fields. Future research should focus on expanding the sample size to refine the categorization and comparison processes across diverse human liver tissue types.

Background: Understanding the changes occurring in biological tissue during thermal ablation is at the heart of many current challenges in both therapy and medical imaging research. Purpose: The objective of this work is to quantitatively interpret the scattering response of human liver samples, before and after thermal ablation. We report acoustic measurements performed involving n = 21 human liver samples. Thermal ablation is achieved at temperatures between 45 and 80°C and quantification of the irreversible changes in acoustic attenuation and Backscattering Coefficient (BSC) is reported, with a particular attention to the latter. Methods: Both attenuation coefficient and BSCs were measured in the frequency range from 10 to 52 MHz. Scans were performed before heating and after cooling down. Attenuation coefficients were calculated using spectral difference method and BSC estimated using the reference phantom method. Results: Strong increases of attenuation coefficients and BSCs with heating temperature were observed. Quantitative ultrasonic parameters obtained with the polydisperse structure factor model (poly‐SFM)are compared to histological observations and seen to be close to hepatocyte mean diameter (HMD). Conclusions: The results presented in this study provide a description of the impact of thermal ablation in human liver tissue on acoustic attenuation and the BSC. For the first time, quantitative agreement between the Effective Scatterer Diameter (ESD) estimated from BSC and HMD was shown, highlighting the important role of cellular network in the scattering response of the medium. This core result is an important step toward the determination of the nature of scattering sources in biological tissues. [ABSTRACT FROM AUTHOR]

Liver injury is one of the most severe traffic trauma. The development of accurate liver finite element model is beneficial for improving the biofidelity and validity in crash simulations, aiming to analyze the injury in accidents. 12 human liver finite element models in the current research were constructed from high resolution CT data of a Chinese male 50th percentile human subject, including the main structures like left lobe, right lobe, capsule, parenchyma and falciform ligament. The simulations based on Nava et al. experiment were conducted to validate the models and make comparisons of modeling method accuracy. The results demonstrated that the larger deviation happened to the tetra models due to the stiffer algorithm compared to the hex models, which were more sensitive to element size. The existence of capsule had significant effects on the liver mechanical responses, reducing the liver tissue pressure. Shell elements were more suitable for modeling the capsule. [ABSTRACT FROM AUTHOR]

This paper will introduce novel techniques for a fractional-order model of the human liver involving the Atangana–Baleanu, Atangana–Toufik, and the Fractal fractional method with the nonsingular kernel. These techniques give more accurate and appropriate results. Existence and uniqueness have been developed with the help of fixed-point theory results. Numerical simulations are done from the developed algorithm of the model to elaborate the effect of fractional values and justify the theoretical results. Such kind of analysis will be useful for further investigation of epidemic diseases, and also provide a better understanding of disease dynamics to overcome the effect of disease in a community. [ABSTRACT FROM AUTHOR]

Acute-on-chronic liver failure (ACLF) is a major cause of liver-related death worldwide, but its key pathological features remain incompletely defined. This study aimed to reveal the molecular basis of hepatitis B virus-related ACLF (HBV-ACLF) by transcriptome sequencing of human liver tissue. A total of 18 human liver tissues from patients with different stages of HBV-related disease were collected for RNA sequencing, and liver tissues from patients and mouse models with ACLF were used for subsequent validation. Specifically, 6,853 differentially expressed genes (DEGs) and 5,038 differentially expressed transcripts were identified in patients with ACLF compared to patients with chronic hepatitis B (CHB) and normal controls (NCs). Investigation of functional by KEGG pathway enrichment analysis revealed prominent immune and metabolic dysregulation at the ACLF stage. We found that the key genes FGF19, ADCY8 and KRT17, which are related to immunometabolic disturbances, were significantly upregulated in the progression of ACLF. The three key genes were validated in human and mouse samples, indicating their prognostic and therapeutic potential in ACLF. In summary, our work reveals that immunometabolic disorder is involved in HBV-ACLF pathogenesis and indicates that FGF19, ADCY8 and KRT17 may be sensitive biomarkers for HBV-related ACLF. [ABSTRACT FROM AUTHOR]

IL-17A is considered to guide liver inflammation and fibrosis. From twenty-two human liver samples of different fibrosis stages (F0 to F4), IL-17A, IL-22, and TGFβ1 protein expression in liver tissue lysates were analyzed. Ten paired samples of liver tissue (F0–F1 stage) and blood from the same patient were used to analyze intrahepatic and blood T-lymphoid IL-17A+ cells by flow cytometry. The analyses have been performed regardless of pathology, considering the stage of fibrosis. Human liver tissue was used for the primary human liver slice cultures, followed by subsequent cytokine stimulation and fibrotic markers' analysis by ELISA. IL-17A production in human liver tissue was significantly higher in the early fibrotic stage compared with the advanced stage. Th17 T cells and, to a lesser extent, MAIT cells were the main sources of IL-17A in both compartments, the liver and the blood. Moreover, the presence of liver Th17IL-17A+INFγ+ cells was detected in the liver. IL-17A stimulation of human liver slice culture increased the expression of profibrotic and pro-inflammatory markers. IL-17A, secreted by Th17 and MAIT cells in the liver, triggered fibrosis by inducing the expression of IL-6 and profibrotic markers and could be a target for antifibrotic treatment. Further amplitude studies are needed to confirm the current results. [ABSTRACT FROM AUTHOR]

2-Methylfuran (2-MF) is a well-known industrial chemical and also formed via thermal treatment of food. One main source of 2-MF in the human diet is coffee. 2-MF is known to form 3-acetylacrolein (AcA, 4-oxopent-2-enal) via cytochrome P 450 metabolism and further reacts with amino acids in vivo. Still the reactivity toward other biomolecules is rather scarce. Therefore, AcA was synthesized, and its reaction with 2'-deoxyadenosine (dA), 2'deoxyguanosine (dG), 2'deoxycytosine (dC), and 2'-deoxythymidine (dT) was tested. For this purpose, adduct formation was performed by acid hydrolysis of 2,5-dihydro-2,5-dimethoxy-2-methylfuran (DHDMMF) as well as pure AcA. The structures of these adducts were confirmed by UPLC-ESI + -MS/MS fragmentation patterns and 1 H-/ 13 CNMR spectra. Except for dT, which showed no reactivity, all adducts of AcA were characterized, which enabled the development of sensitive quantification methods via (U)HPLC-ESI ± -MS/MS. Pure AcA was synthesized by oxidation of 2-MF using dimethyldioxirane (DMDO), and its behavior in aqueous medium was studied. Incubations of AcA and isolated DNA of primary rat hepatocytes (pRH) showed time- and dose-dependent formation of the identified DNA adducts dA-AcA, dG-AcA, or dC-AcA. In contrast, the DNA adducts dA-AcA, dG-AcA, or dC-AcA were not detected on a cellular level when pRH were incubated with 2-MF or AcA. This indicates an efficient detoxification or reaction with biomolecules in the cell, although the induction of other DNA damage, possibly also by other metabolites, cannot be ruled out in principle.

In this work, we aim to propose a mathematical formalism for the human liver involving Caputo fractional derivative (CFD) with modified parameters. Regarding the proposed fractional order model (FOM), the positivity and boundedness theory of an explicit solution are established by using the fractional mean-value theorem in the sense of Caputo. In addition, the validation of the FOM is provided to ensure that it complies with the medical interpretation and we also discuss the stability of unique equilibrium point (EP) for the presented model. We apply the Generalized Mittag–Leffler function method (GMLFM) and Predictor–Corrector method (PCM) to solve this model. The GMLFM is used to give the proper results as a series of the time variable to obtain the desired solution at any time and the PCM is implemented to obtain the solution of the FOM over a long time period. The tabled results from this study are compared with the real clinical data and with other methods. The simulations approximate the real clinical data and support the obtained theoretical results. Precisely, the outcomes of the FOM by using the proposed methods coincide with most of the clinical data that means it more accurate. Ultimately, the graphical results indicate the superiority of the proposed FOM over the classical model.

Patients with acute-on-chronic liver failure (ACLF) are at risk of developing acute hepatic decompensation and organ failures with an unraveled complex mechanism. An altered immune response toward insults in cirrhotic compared with healthy livers may contribute to the ACLF development. Therefore, we aim to investigate the differences in inflammatory responses between cirrhotic and healthy livers using human precision-cut liver slices (PCLSs) upon the lipopolysaccharide (LPS) challenge. PCLSs prepared from livers of patients with cirrhosis or healthy donors of liver transplantation were incubated ex vivo with or without LPS for up to 48 h. Viability test, qRT-PCR, and multiplex cytokine assay were performed. Regulation of the LPS receptors during incubation or with LPS challenge differed between healthy versus cirrhotic PCLSs. LPS upregulated TLR-2 in healthy PCLSs solely (P < 0.01). Culturing for 48 h induced a stronger inflammatory response in the cirrhotic than healthy PCLS. Upon LPS stimulation, cirrhotic PCLSs secreted more proinflammatory cytokines (IL-8, IL-6, TNF-a, eotaxin, and VEGF) significantly and less antiinflammatory cytokine (IL-1ra) than those of healthy. In summary, cirrhotic PCLSs released more proinflammatory and less antiinflammatory cytokines after LPS stimuli than healthy, leading to dysregulated inflammatory response. These events could possibly resemble the liver immune response in ACLF. [ABSTRACT FROM AUTHOR]

Animal models and clinical studies suggest an influence of angiotensin II (AngII) on the pathogenesis of liver diseases via the renin–angiotensin system. AngII application increases portal blood pressure, reduces bile flow, and increases permeability of liver tight junctions. Establishing the subcellular localization of angiotensin II receptor type 1 (AT1R), the main AngII receptor, helps to understand the effects of AngII on the liver. We localized AT1R in situ in human and porcine liver and porcine gallbladder by immunohistochemistry. In order to do so, we characterized commercial anti-AT1R antibodies regarding their capability to recognize heterologous human AT1R in immunocytochemistry and on western blots, and to detect AT1R using overlap studies and AT1R-specific blocking peptides. In hepatocytes and canals of Hering, AT1R displayed a tram-track-like distribution, while in cholangiocytes AT1R appeared in a honeycomb-like pattern; i.e., in liver epithelia, AT1R showed an equivalent distribution to that in the apical junctional network, which seals bile canaliculi and bile ducts along the blood–bile barrier. In intrahepatic blood vessels, AT1R was most prominent in the tunica media. We confirmed AT1R localization in situ to the plasma membrane domain, particularly between tight and adherens junctions in both human and porcine hepatocytes, cholangiocytes, and gallbladder epithelial cells using different anti-AT1R antibodies. Localization of AT1R at the junctional complex could explain previously reported AngII effects and predestines AT1R as a transmitter of tight junction permeability. [ABSTRACT FROM AUTHOR]

Adapted fixation methods for electron microscopy allowed us to study liver cell fine structure in 217 biopsies of intact human livers over the course of 10 years. The following novel observations and concepts arose: single fat droplets in parenchymal cells can grow to a volume four times larger than the original cell, thereby extremely marginalizing the cytoplasm with all organelles. Necrosis of single parenchymal cells, still containing one huge fat droplet, suggests death by fat in a process of single-cell steatonecrosis. In a later stage of single-cell steatonecrosis, neutrophils and erythrocytes surround the single fat droplet, forming an inflammatory fat follicle indicating the apparent onset of inflammation. Also, fat droplets frequently incorporate masses of filamentous fragments and other material, most probably representing Mallory substance. No other structure or material was found that could possibly represent Mallory bodies. We regularly observe the extrusion of huge fat droplets, traversing the peripheral cytoplasm of parenchymal cells, the Disse space and the endothelium. These fat droplets fill the sinusoid as a sinusoidal lipid embolus. In conclusion, adapted methods of fixation applied to human liver tissue revealed that single, huge fat droplets cause necrosis and inflammation in single parenchymal cells. Fat droplets also collect Mallory substance and give rise to sinusoidal fat emboli. Therefore, degreasing of the liver seems to be an essential therapeutic first step in the self-repairing of non-alcoholic fatty liver disease. This might directly reduce single-cell steatotic necrosis and inflammation as elements in non-alcoholic steatohepatitis progression. [ABSTRACT FROM AUTHOR]

Pirfenidone is a first-line drug for the treatment of idiopathic pulmonary fibrosis. The primary metabolic pathways of pirfenidone in humans are 5-hydroxylation and subsequent oxidation to 5-carboxylpirfenidone. The aims of this study were to determine the cytochrome P450 isoforms responsible for pirfenidone 5-hydroxylation and to evaluate their contributions in human liver microsomes (HLM). Among the recombinant P450 isoforms, CYP1A2, CYP2D6, CYP2C19, CYP2A6, and CYP2B6 were shown to catalyse the 5-hydroxylation of pirfenidone. Pirfenidone 5-hydroxylase activity by HLM was inhibited by α-naphthoflavone (by 45%), 8-methoxypsolaren (by 84%), tranylcypromine (by 53%), and quinidine (by 15%), which are CYP1A2, CYP1A2/CYP2A6/CYP2C19, CYP2A6/CYP2C19, and CYP2D6 inhibitors, respectively. In 17 individual HLM donors, pirfenidone 5-hydroxylase activity was significantly correlated with phenacetin O-deethylase (r = 0.89, P < 0.001) and S-mephenytoin 4'-hydroxylase activities (r = 0.51, P < 0.05), which are CYP1A2 and CYP2C19 marker activities, respectively. By using the relative activity factors, the contributions of CYP1A2, CYP2C19, and CYP2D6 to pirfenidone 5-hydroxylation in the human liver were 72.8%, 11.8%, and 8.9%, respectively. In conclusion, we clearly demonstrated the predominant P450 involved in pirfenidone 5-hydroxylation in the human liver is CYP1A2, with CYP2C19 and CYP2D6 playing a minor role. [ABSTRACT FROM AUTHOR]

Background & Aims: Unlike protein-coding genes, the majority of human long non-coding RNAs (lncRNAs) lack conservation based on their sequences, posing a challenge for investigating their role in a pathophysiological context for clinical translation. This study explores the hypothesis that non-conserved lncRNAs in human and mouse livers may share similar metabolic functions, giving rise to functionally conserved lncRNA metabolic regulators (fcLMRs)., Methods: We developed a sequence-independent strategy to select putative fcLMRs, and performed extensive analysis to determine the functional similarities of putative human and mouse LMR pairs (h/mLMRs)., Results: We found that several pairs of putative fcLMRs share similar functions in regulating gene expression. We further demonstrated that a pair of fcLMRs, h/mLMR1, robustly regulated triglyceride levels by modulating the expression of a similar set of lipogenic genes. Mechanistically, h/mLMR1 binds to PABPC1, a regulator of protein translation, via short motifs on either lncRNA with divergent sequences but similar structures. This interaction inhibits protein translation, activating an amino acid-mTOR-SREBP1 axis to regulate lipogenic gene expression. Intriguingly, PABPC1-binding motifs on each lncRNA fully rescued the functions of their corresponding LMRs in the opposite species. Given the elevated expression of h/mLMR1 in humans and mice with hepatic steatosis, the PABPC1-binding motif on hLMR1 emerges as a potential non-conserved human drug target whose functions can be fully validated in a physiologically relevant setting before clinical studies., Conclusions: Our study supports that fcLMRs represent a novel and prevalent biological phenomenon, and deep phenotyping of genetic mLMR mouse models constitutes a powerful approach to understand the pathophysiological role of lncRNAs in the human liver., Competing Interests: Disclosures The authors have declared that no conflict of interest exists.

In this work, we aim to propose a mathematical formalism for the human liver involving Caputo fractional derivative (CFD) with modified parameters. Regarding the proposed fractional order model (FOM), the positivity and boundedness theory of an explicit solution are established by using the fractional mean-value theorem in the sense of Caputo. In addition, the validation of the FOM is provided to ensure that it complies with the medical interpretation and we also discuss the stability of unique equilibrium point (EP) for the presented model. We apply the Generalized Mittag–Leffler function method (GMLFM) and Predictor–Corrector method (PCM) to solve this model. The GMLFM is used to give the proper results as a series of the time variable to obtain the desired solution at any time and the PCM is implemented to obtain the solution of the FOM over a long time period. The tabled results from this study are compared with the real clinical data and with other methods. The simulations approximate the real clinical data and support the obtained theoretical results. Precisely, the outcomes of the FOM by using the proposed methods coincide with most of the clinical data that means it more accurate. Ultimately, the graphical results indicate the superiority of the proposed FOM over the classical model. [ABSTRACT FROM AUTHOR]

The study of the function of vital organs of the body by mathematical models is one of the most important and interesting topics for researchers. In this work, we intend to study the mathematical model of human liver function using the fractional order derivative with Caputo approach. The Adomian Analytical Analysis (ADM) method will be used to solve the system of fractional-order differential equations obtained in the new model of the liver. We also provide a numerical simulation for the results obtained from the fractional order system and the integer order system using the available clinical data. [ABSTRACT FROM AUTHOR]

Nine forms of recombinant cytochrome P450 (P450 or CYP) enzymes were used to study roles of individual P450 enzymes in the oxidation of flavone and some other flavonoids, 4′-hydroxyflavone and 4′-, 3′-, and 2′-methoxyflavones, by human liver microsomes using LC-MS/MS analysis. As has been reported previously , 4′-, 3′-, and 2′-methoxyflavones were preferentially O-demethylated by human liver P450 enzymes to form 4′-, 3′-, and 2′-hydroxylated flavones and also 3′,4′-dihydroxyflavone from the former two substrates. In comparisons of product formation by oxidation of these methoxylated flavones, CYP2A6 was found to be a major enzyme catalysing flavone 4′- and 3′-hydroxylations by human liver microsomes but did not play significant roles in 2′-hydroxylation of flavone, O-demethylations of three methoxylated flavones, and the oxidation of 4′-hydroxyflavone to 3′,4′-dihydroxyflavone. The effects of anti-CYP2A6 IgG and chemical P450 inhibitors suggested that different P450 enzymes, as well as CYP2A6, catalysed oxidation of these flavonoids at different positions by liver microsomes. These studies suggest that CYP2A6 catalyses flavone 4′- and 3′-hydroxylations in human liver microsomes and that other P450 enzymes have different roles in oxidizing these flavonoids. [ABSTRACT FROM AUTHOR]

This study was conducted to compare the anatomical histological and histochemical structure of human liver with the liver of domestic rabbits to evaluate the quality of liver of rabbit to be used as a model of mammalian species in the experimental researches. Ten liver biopsies of human (Group I) and same number of liver specimens of domestic rabbit (Group II) were used in this study. The liver specimens were processed and stained with Harris Hematoxylin and Eosin stain and Masson's Trichrome stain and examined under light microscope. For histochemical examination, Periodic Acid Schiff's, Best's carmine and Gomori's Alkaline Phosphatase stains were used. Histomorphometric measurements were done to compare some parameters between the two groups. Transabdominal ultrasound examination was done to compare ultrasonic appearance of the liver in both groups. The work was done from May 2018 to December 2018. The study revealed more lobulated liver of domestic rabbits, hexagonal lobules are mingled with each other in rabbit's liver compared to human, larger hepatocytes, with higher percentage of binucleated hepatocytes and thicker capsule in the human's liver than in the rabbit's liver. Histochemical results revealed less carbohydrates, less glycogen content with less activity of alkaline phosphatase in the cytoplasm of hepatocytes of the human's liver. The ultrasound image revealed denser hyperechoic diaphragm and fibrous capsule, non-homogenous low echogenicity of parenchyma of the liver of human compared to the rabbit's liver. We conclude that the basic anatomical and histological structure is more or less similar in the human's liver andliver of domestic rabbit but the few differences might be related to the evolutional variations among mammals thus the value of the liver tissue of domestic rabbit to be used in the experimental researches is significant.

We report the deformation of the needle during its insertion into the human liver in the context of surgery simulation of the high-robotic-assisted intraoperative treatment of liver tumors based on the integrated imaging-molecular diagnosis. The bee needle is modeled as a flexible thread within the framework of the nonlinear elasticity theory. The motion equations of the needle are similar to Tzitzeica equations of surfaces which are invariant under the group of centro-affine transformations. That means the surfaces tend to minimize their area and have a minimal Dirichlet energy of how variable a function is. The closed form solutions are obtained for deformation of the needle. In addition, the collision between the needle and the tissues is modeled as a minimization problem. [ABSTRACT FROM AUTHOR]

ABC transporters (ATP‐binding cassette transporter) traffic drugs and their metabolites across membranes, making ABC transporter expression levels a key factor regulating local drug concentrations in different tissues and individuals. Yet, quantification of ABC transporters remains challenging because they are large and low‐abundance transmembrane proteins. Here, we analysed 200 samples of crude and membrane‐enriched fractions from human liver, kidney, intestine, brain microvessels and skin, by label‐free quantitative mass spectrometry. We identified 32 (out of 48) ABC transporters: ABCD3 was the most abundant in liver, whereas ABCA8, ABCB2/TAP1 and ABCE1 were detected in all tissues. Interestingly, this atlas unveiled that ABCB2/TAP1 may have TAP2‐independent functions in the brain and that biliary atresia (BA) and control livers have quite different ABC transporter profiles. We propose that meaningful biological information can be derived from a direct comparison of these data sets. [ABSTRACT FROM AUTHOR]

Abstract Background Titanium dioxide (TiO2) is produced at high volumes and applied in many consumer and food products. Recent toxicokinetic modelling indicated the potential of TiO2 to accumulate in human liver and spleen upon daily oral exposure, which is not routinely investigated in chronic animal studies. A health risk from nanosized TiO2 particle consumption could not be excluded then. Results Here we show the first quantification of both total titanium (Ti) and TiO2 particles in 15 post-mortem human livers and spleens. These low-level analyses were enabled by the use of fully validated (single particle) inductively coupled plasma high resolution mass spectrometry ((sp)ICP-HRMS) detection methods for total Ti and TiO2 particles. The presence of TiO2 in the particles in tissues was confirmed by Scanning Electron Microscopy with energy dispersive X-ray spectrometry. Conclusions These results prove that TiO2 particles are present in human liver and spleen, with ≥24% of nanosize (

Abstract Introduction Murine hepatic NK cells exhibit adaptive features, with liver‐specific adhesion molecules CXCR6 and CD49a acting as surface markers. Methods We investigated human liver‐resident CXCR6+ and CD49a+ NK cells using RNA sequencing, flow cytometry, and functional analysis. We further assessed the role of cytokines in generating NK cells with these phenotypes from the peripheral blood. Results Hepatic CD49a+ NK cells could be induced using cytokines and produce high quantities of IFNγ and TNFα, in contrast to hepatic CXCR6+ NK cells. RNA sequencing of liver‐resident CXCR6+ NK cells confirmed a tolerant immature phenotype with reduced expression of markers associated with maturity and cytotoxicity. Liver‐resident double‐positive CXCR6 + CD49a+ hepatic NK cells are immature but maintain high expression of Th1 cytokines as observed for single‐positive CD49a+ NK cells. We show that stimulation with activating cytokines can readily induce upregulation of both CD49a and CXCR6 on NK cells in the peripheral blood. In particular, IL‐12 and IL‐15 can generate CXCR6 + CD49a+ NK cells in vitro from NK cells isolated from the peripheral blood, with comparable phenotypic and functional features to liver‐resident CD49a+ NK cells, including enhanced IFNγ and NKG2C expression. Conclusion IL‐12 and IL‐15 may be key for generating NK cells with a tissue‐homing phenotype and strong Th1 cytokine profile in the blood, and links peripheral activation of NK cells with tissue‐homing. These findings may have important therapeutic implications for immunotherapy of chronic liver disease.

Aims/hypothesis: IRS2 is an important molecular switch that mediates insulin signalling in the liver. IRS2 dysregulation is responsible for the phenomenon of selective insulin resistance that is observed in type 2 diabetes. We hypothesise that epigenetic mechanisms are involved in the regulation of IRS2 in the liver of obese and type 2 diabetic individuals. Methods: DNA methylation of seven CpG sites was studied by bisulphite pyrosequencing and mRNA and microRNA (miRNA) expression was assessed by quantitative real-time PCR in liver biopsies of 50 obese non-diabetic and 31 obese type 2 diabetic participants, in a cross-sectional setting. Methylation-sensitive luciferase assays and electrophoretic mobility shift assays were performed. Furthermore, HepG2 cells were treated with insulin and high glucose concentrations to induce miRNA expression and IRS2 downregulation. Results: We found a significant downregulation of IRS2 expression in the liver of obese individuals with type 2 diabetes (0.84 ± 0.08-fold change; p = 0.0833; adjusted p value [pa] = 0.0417; n = 31) in comparison with non-diabetic obese participants (n = 50). This downregulation correlated with hepatic IRS2 DNA methylation at CpG5. Additionally, CpG6, which is located in intron 1 of IRS2, was hypomethylated in type 2 diabetes; this site spans the sterol regulatory element binding transcription factor 1 (SREBF1) recognition motif, which likely acts as transcriptional repressor. The adjacent polymorphism rs4547213 (G>A) was significantly associated with DNA methylation at a specificity-protein-1 (SP1) binding site (CpG3). Moreover, DNA methylation of cg25924746, a CpG site located in the shore region of the IRS2 promoter-associated CpG island, was increased in the liver of individuals with type 2 diabetes, as compared with those without diabetes. A second epigenetic mechanism, upregulation of hepatic miRNA hsa-let-7e-5p (let-7e-5p) in obese individuals with type 2 diabetes (n = 29) vs non-diabetic obese individuals (n = 49) (1.2 ± 0.08-fold change; p = 0.0332; pa = 0.0450), is likely to act synergistically with altered IRS2 DNA methylation to decrease IRS2 expression. Mechanistic in vitro experiments demonstrated an acute upregulation of let-7e-5p expression and simultaneous IRS2 downregulation in a liver (HepG2) cell line upon hyperinsulinaemic and hyperglycaemic conditions. Conclusions/interpretation: Our study highlights a new multi-layered epigenetic network that could be involved in subtle dysregulation of IRS2 in the liver of individuals with type 2 diabetes. This might lead to fine-tuning of IRS2 expression and is likely to be supplementary to the already known factors regulating IRS2 expression. Thereby, our findings could support the discovery of new diagnostic and therapeutic strategies for type 2 diabetes. [ABSTRACT FROM AUTHOR]

Human liver CD69+CD8+ T cells are ~95% CD103- and ~5% CD103+. Although CD69+CD103+CD8+ T cells show tissue residency and robustly respond to antigens, CD69+CD103-CD8+ T cells are not yet well understood. Liver perfusate and paired peripheral blood were collected from healthy living donors and recipients with cirrhosis during liver transplantation. Liver tissues were obtained from patients with acute hepatitis A. Phenotypic and functional analyses were performed by flow cytometry. Gene expression profiles were determined by microarray and quantitative reverse transcription PCR. PT-2385 was used to inhibit hypoxia-inducible factor (HIF)-2α. Human liver CD69+CD103-CD8+ T cells exhibited HIF-2α upregulation with a phenotype of tissue residency and terminal differentiation. CD103- cells comprised non-hepatotropic virus-specific T cells as well as hepatotropic virus-specific T cells, but CD103+ cells exhibited only hepatotropic virus specificity. Although CD103- cells were weaker effectors on a per cell basis than CD103+ cells, following T cell receptor or interleukin-15 stimulation, they remained the major CD69+CD8+ effector population in the liver, surviving with less cell death. An HIF-2α inhibitor suppressed the effector functions and survival of CD69+CD103-CD8+ T cells. In addition, HIF-2α expression in liver CD69+CD103-CD8+ T cells was significantly increased in patients with acute hepatitis A or cirrhosis. Liver CD69+CD103-CD8+ T cells are tissue resident and terminally differentiated, and their effector functions depend on HIF-2α. Furthermore, activation of liver CD69+CD103-CD8+ T cells with HIF-2α upregulation is observed during liver pathology. The immunologic characteristics and the role of CD69+CD103-CD8+ T cells, which are a major population of human liver CD8+ T cells, remain unknown. Our study shows that these T cells have a terminally differentiated tissue-resident phenotype, and their effector functions depend on a transcription factor, HIF-2α. Furthermore, these T cells were activated and expressed higher levels of HIF-2α in liver pathologies, suggesting that they play an important role in immune responses in liver tissues and the pathogenesis of human liver disease. • CD69+CD103-CD8+ T cells are the major population of human liver CD8+ T cells. • Human liver CD69+CD103-CD8+ T cells have a terminally differentiated and tissue-resident phenotype. • Human liver CD69+CD103-CD8+ T cells specifically upregulate HIF-2α. • Effector functions and survival of human liver CD69+CD103-CD8+ T cells depend on HIF-2α. • Activation of liver CD69+CD103-CD8+ T cells with HIF-2α upregulation is observed during acute and chronic liver pathology. [ABSTRACT FROM AUTHOR]

The liver is the largest solid organ and a primary metabolic hub. In recent years, intact cell nuclei were used to perform single-nuclei RNA-seq (snRNA-seq) for tissues difficult to dissociate and for flash-frozen archived tissue samples to discover unknown and rare cell subpopulations. In this study, we performed snRNA-seq of a liver sample to identify subpopulations of cells based on nuclear transcriptomics. In 4282 single nuclei, we detected, on average, 1377 active genes and we identified seven major cell types. We integrated data from 94,286 distal interactions (p < 0.05) for 7682 promoters from a targeted chromosome conformation capture technique (HiCap) and mass spectrometry proteomics for the same liver sample. We observed a reasonable correlation between proteomics and in silico bulk snRNA-seq (r = 0.47) using tissue-independent gene-specific protein abundancy estimation factors. We specifically looked at genes of medical importance. The DPYD gene is involved in the pharmacogenetics of fluoropyrimidine toxicity and some of its variants are analyzed for clinical purposes. We identified a new putative polymorphic regulatory element, which may contribute to variation in toxicity. Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and we investigated all known risk genes. We identified a complex regulatory landscape for the SLC2A2 gene with 16 candidate enhancers. Three of them harbor somatic motif breaking and other mutations in HCC in the Pan Cancer Analysis of Whole Genomes dataset and are candidates to contribute to malignancy. Our results highlight the potential of a multi-omics approach in the study of human diseases. [ABSTRACT FROM AUTHOR]

Abstract Background Recent advances in omics technology have produced a large amount of liver-related data. A comprehensive and up-to-date source of liver-related data is needed to allow biologists to access the latest data. However, current liver-related data sources each cover only a specific part of the liver. It is difficult for them to keep pace with the rapid increase of liver-related data available at those data resources. Integrating diverse liver-related data is a critical yet formidable challenge, as it requires sustained human effort. Results We present LiverWiki, a first wiki-based database that integrates liver-related genes, homolog genes, gene expressions in microarray datasets and RNA-Seq datasets, proteins, protein interactions, post-translational modifications, associated pathways, diseases, metabolites identified in the metabolomics datasets, and literatures into an easily accessible and searchable resource for community-driven sharing. LiverWiki houses information in a total of 141,897 content pages, including 19,787 liver-related gene pages, 17,077 homolog gene pages, 50,251 liver-related protein pages, 36,122 gene expression pages, 2067 metabolites identified in the metabolomics datasets, 16,366 disease-related molecules, and 227 liver disease pages. Other than assisting users in searching, browsing, reviewing, refining the contents on LiverWiki, the most important contribution of LiverWiki is to allow the community to create and update biological data of liver in visible and editable tables. This integrates newly produced data with existing knowledge. Implemented in mediawiki, LiverWiki provides powerful extensions to support community contributions. Conclusions The main goal of LiverWiki is to provide the research community with comprehensive liver-related data, as well as to allow the research community to share their liver-related data flexibly and efficiently. It also enables rapid sharing new discoveries by allowing the discoveries to be integrated and shared immediately, rather than relying on expert curators. The database is available online at http://liverwiki.hupo.org.cn /.

Macrophages play central roles in inflammatory reactions and initiation of immune responses during infections. More than 80% of total tissue macrophages are described to be located in the liver as liver-resident macrophages, also named Kupffer cells (KCs). While studies in mice have established a central role of liver-resident KCs in regulating liver inflammation, their phenotype and function are not well-characterized in humans. Comparing paired human liver and peripheral blood samples, we observed significant differences in the distribution of macrophage (Mφ) subsets, with lower frequencies of CD14hiCD16lo and higher frequencies of CD14int−hiCD16int Mφ in human livers. Intrahepatic Mφ consisted of diverse subsets with differential expression of CD49a, a liver-residency marker previously described for human and mice NK cells, and VSIG4 and/or MARCO, two recently described human tissue Mφ markers. Furthermore, intrahepatic CD49a+ Mφ expressed significantly higher levels of maturation and activation markers, exhibited higher baseline levels of TNF-α, IL-12, and IL-10 production, but responded less to additional in vitro TLR stimulation. In contrast, intrahepatic CD49a− Mφ were highly responsive to stimulation with TLR ligands, similar to what was observed for CD49a− monocytes (MOs) in peripheral blood. Taken together, these studies identified populations of CD49a+, VSIG4+, and/or MARCO+ Mφ in human livers, and demonstrated that intrahepatic CD49a+ Mφ differed in phenotype and function from intrahepatic CD49a− Mφ as well as from peripheral blood-derived monocytes.

T-cell immunity in the liver is tightly regulated to prevent chronic liver inflammation in response to antigens and toxins derived from food and intestinal bacterial flora. Since the main sites of T cell activation in response to foreign components entering solid tissues are the draining lymph nodes (LN), we aimed to study whether Antigen-Presenting Cell (APC) subsets in human liver lymph-draining LN show features that may contribute to the immunologically tolerant liver environment. Healthy liver LN, iliac LN, spleen and liver perfusates were obtained from multi-organ donors, while diseased liver LN were collected from explanted patient livers. Inguinal LN were obtained from kidney transplant recipients. Mononuclear cells were isolated from fresh tissues, and immunophenotypic and functional characteristics of APC subsets were studied using flowcytometry and in ex vivo cultures. Healthy liver-draining LN contained significantly lower relative numbers of CD1c+ conventional dendritic cells (cDC2), plasmacytoid DC (PDC), and CD14+CD163+DC-SIGN+ macrophages (MF) compared to inguinal LN. Compared to spleen, both types of LN contained low relative numbers of CD141hi cDC1. Both cDC subsets in liver LN showed a more activated/mature immunophenotype than those in inguinal LN, iliacal LN, spleen and liver tissue. Despite their more mature status, cDC2 isolated from hepatic LN displayed similar cytokine production capacity (IL-10, IL-12, and IL-6) and allogeneic T cell stimulatory capacity as their counterparts from spleen. Liver LN from patients with inflammatory liver diseases showed a further reduction of cDC1, but had increased relative numbers of PDC and MF. In steady state conditions human liver LN contain relatively low numbers of cDC2, PDC, and macrophages, and relative numbers of cDC1 in liver LN decline during liver inflammation. The paucity of cDC in liver LN may contribute to immune tolerance in the liver environment.

Neural regulation of hepatic metabolism has long been recognized. However, the detailed afferent and efferent innervation of the human liver has not been systematically characterized. This is largely due to the liver's high lipid and pigment contents, causing false-negative (light scattering and absorption) and false-positive (autofluorescence) results in in-depth fluorescence imaging. Here, to avoid the artifacts in three-dimensional (3-D) liver neurohistology, we embed the bleached human liver in the high-refractive-index polymer for tissue clearing and antifade 3-D/Airyscan super-resolution imaging. Importantly, using the paired substance P (SP, sensory marker) and PGP9.5 (pan-neuronal marker) labeling, we detect the sensory nerves in the portal space, featuring the SP + varicosities in the PGP9.5 + nerve bundles/fibers, confirming the afferent liver innervation. Also, using the tyrosine hydroxylase (TH, sympathetic marker) labeling, we identify 1 ) condensed TH + sympathetic nerves in the portal space, 2 ) extension of sympathetic nerves from the portal to the intralobular space, in which the TH + nerve density is 2.6 ± 0.7-fold higher than that of the intralobular space in the human pancreas, and 3 ) the TH + nerve fibers and varicosities contacting the ballooning cells, implicating potential sympathetic influence on hepatocytes with macrovesicular fatty change. Finally, using the vesicular acetylcholine transporter (VAChT, parasympathetic marker), PGP9.5, and CK19 (epithelial marker) labeling with panoramic-to-Airyscan super-resolution imaging, we detect and confirm the parasympathetic innervation of the septal bile duct. Overall, our labeling and 3-D/Airyscan imaging approach reveal the hepatic sensory (afferent) and sympathetic and parasympathetic (efferent) innervation, establishing a clinically related setting for high-resolution 3-D liver neurohistology. NEW & NOTEWORTHY We embed the human liver (vs. pancreas, positive control) in the high-refractive-index polymer for tissue clearing and antifade 3-D/Airyscan super-resolution neurohistology. The pancreas-liver comparison reveals: 1 ) sensory nerves in the hepatoportal space; 2 ) intralobular sympathetic innervation, including the nerve fibers and varicosities contacting the ballooning hepatocytes; and 3 ) parasympathetic innervation of the septal bile duct. Our results highlight the sensitivity and resolving power of 3-D/Airyscan super-resolution imaging in human liver neurohistology.

Liver macrophages (LMs) play a central role in acute and chronic liver pathologies. Investigation of these processes in humans as well as the development of diagnostic tools and new therapeutic strategies require in vitro models that closely resemble the in vivo situation. In our study, we sought to gain further insight into the role of LMs in different liver pathologies and into their characteristics after isolation from liver tissue. For this purpose, LMs were characterized in human liver tissue sections using immunohistochemistry and bioinformatic image analysis. Isolated cells were characterized in suspension using FACS analyses and in culture using immunofluorescence staining and laser scanning microscopy as well as functional assays. The majority of our investigated liver tissues were characterized by anti-inflammatory LMs which showed a homogeneous distribution and increased cell numbers in correlation with chronic liver injuries. In contrast, pro-inflammatory LMs appeared as temporary and locally restricted reactions. Detailed characterization of isolated macrophages revealed a complex disease dependent pattern of LMs consisting of pro- and anti-inflammatory macrophages of different origins, regulatory macrophages and monocytes. Our study showed that in most cases the macrophage pattern can be transferred in adherent cultures. The observed exceptions were restricted to LMs with pro-inflammatory characteristics.

Macrophages play central roles in inflammatory reactions and initiation of immune responses during infections. More than 80% of total tissue macrophages are described to be located in the liver as liver-resident macrophages, also named Kupffer cells (KCs). While studies in mice have established a central role of liver-resident KCs in regulating liver inflammation, their phenotype and function are not well-characterized in humans. Comparing paired human liver and peripheral blood samples, we observed significant differences in the distribution of macrophage (Mφ) subsets, with lower frequencies of CD14hiCD16lo and higher frequencies of CD14int−hiCD16int Mφ in human livers. Intrahepatic Mφ consisted of diverse subsets with differential expression of CD49a, a liver-residency marker previously described for human and mice NK cells, and VSIG4 and/or MARCO, two recently described human tissue Mφ markers. Furthermore, intrahepatic CD49a+ Mφ expressed significantly higher levels of maturation and activation markers, exhibited higher baseline levels of TNF-α, IL-12, and IL-10 production, but responded less to additional in vitro TLR stimulation. In contrast, intrahepatic CD49a− Mφ were highly responsive to stimulation with TLR ligands, similar to what was observed for CD49a− monocytes (MOs) in peripheral blood. Taken together, these studies identified populations of CD49a+, VSIG4+, and/or MARCO+ Mφ in human livers, and demonstrated that intrahepatic CD49a+ Mφ differed in phenotype and function from intrahepatic CD49a− Mφ as well as from peripheral blood-derived monocytes. [ABSTRACT FROM AUTHOR]

Thymidine phosphorylase (TP; EC 2.4.2.4) catalyzes the reversible phosphorolysis of thymidine, deoxyuridine, and their analogues to their respective nucleobases and 2-deoxy-α- d -ribose-1-phosphate (dRib-1- P). TP is a key enzyme in the pyrimidine salvage pathways. Activity of the enzyme is crucial in angiogenesis, cancer chemotherapy, radiotherapy, and tumor imaging, Nevertheless, a complete set of kinetic parameters has never been reported for any human TP. This study describes the kinetic mechanism and regulation of native human hepatic TP. The liver is a main site of pyrimidine metabolism and contains high levels of TP. Initial velocity and product inhibition studies demonstrated that the basic mechanism of this enzyme is a sequential random bi-bi mechanism. Initial velocity studies showed an intersecting pattern, consistent with substrate-enzyme-co-substrate complex formation, and a binding pattern indicating that the binding of the substrate interferes with the binding of the co-substrate and vice versa. Estimated kinetic parameters were K Thymidine = 284 ± 55, K P i = 5.8 ± 1.9, K Thymine = 244 ± 69, and K dRib-1- P = 90 ± 33 μM. Thymine was a product activator, but becomes a substrate inhibitor at concentrations eight times higher than its K m. dRib-1- P was a non-competitive product inhibitor of the forward reaction. It bounded better to the Enzyme● P i complex than the free enzyme, but had better affinity to the free enzyme than the Enzyme●Thymidine complex. In the reverse reaction, dRib-1- P enhanced the binding of thymine. The enhancement of the thymine binding along with the fact that dRib-1- P was a non-competitive product inhibitor suggests the presence of another binding site for dRib-1- P on the enzyme. [ABSTRACT FROM AUTHOR]

Detection of fatty liver disease (steatosis) from the ultra sound (US) images using pattern recognition techniques is attempted by many authors. Different pre-processing methods, feature extraction, feature selection and classification models are reported in the literature. The present work uses a hitherto unexplored property of liver texture. A careful visual observation reveals that the liver texture is anisotropic. A novel set of features exploiting this anisotropy is explicitly proposed. These anisotropy features are derived from gray level difference histogram (GLDH), pair correlation function (PCF), probabilistic local directionality statistics and randomness of texture (GLCM-χ8). For comparison with other features, the most popular gray level co-occurrence matrix (GLCM) derived features are also extracted. Accordingly, three alternative data sets are prepared to classify the images with five different classifiers –Bayesian, multilayer perceptron (MLP), probabilistic neural network (PNN), learning vector quantisation (LVQ) and support vector machine (SVM). A comparative evaluation in terms of specificity, sensitivity, discrimination score and accuracy has been made while classifying US images of human livers. On the basis of results this paper enumerates as to how the anisotropy feature provides better entity for classification purpose in the present context. It is also shown that the highest accuracy of 99% is obtained using anisotropy features with PNN. Anisotropy features leads to 100% sensitivity with PNN and SVM. The present classification system with anisotropy features outperforms the existing models available in the literature keeping in mind the simplicity of the algorithm. [ABSTRACT FROM AUTHOR]

T-cell immunity in the liver is tightly regulated to prevent chronic liver inflammation in response to antigens and toxins derived from food and intestinal bacterial flora. Since the main sites of T cell activation in response to foreign components entering solid tissues are the draining lymph nodes (LN), we aimed to study whether Antigen-Presenting Cell (APC) subsets in human liver lymph-draining LN show features that may contribute to the immunologically tolerant liver environment. Healthy liver LN, iliac LN, spleen and liver perfusates were obtained from multi-organ donors, while diseased liver LN were collected from explanted patient livers. Inguinal LN were obtained from kidney transplant recipients. Mononuclear cells were isolated from fresh tissues, and immunophenotypic and functional characteristics of APC subsets were studied using flowcytometry and in ex vivo cultures. Healthy liver-draining LN contained significantly lower relative numbers of CD1c+ conventional dendritic cells (cDC2), plasmacytoid DC (PDC), and CD14+CD163+DC-SIGN+ macrophages (MF) compared to inguinal LN. Compared to spleen, both types of LN contained low relative numbers of CD141hi cDC1. Both cDC subsets in liver LN showed a more activated/mature immunophenotype than those in inguinal LN, iliacal LN, spleen and liver tissue. Despite their more mature status, cDC2 isolated from hepatic LN displayed similar cytokine production capacity (IL-10, IL-12, and IL-6) and allogeneic T cell stimulatory capacity as their counterparts from spleen. Liver LN from patients with inflammatory liver diseases showed a further reduction of cDC1, but had increased relative numbers of PDC and MF. In steady state conditions human liver LN contain relatively low numbers of cDC2, PDC, and macrophages, and relative numbers of cDC1 in liver LN decline during liver inflammation. The paucity of cDC in liver LN may contribute to immune tolerance in the liver environment. [ABSTRACT FROM AUTHOR]

Highlights • A reduced order model (ROM) encompassing the anatomical liver shape is created. • The ROM is able to reproduce the mechanical behavior with high fidelity. • The solution is computed in a 11 dimensional subspace. Abstract Background and objective This paper focuses on computer simulation aspects of Digital Twin models in the medical framework. In particular, it addresses the need of fast and accurate simulators for the mechanical response at tissue and organ scale and the capability of integrating patient-specific anatomy from medical images to pinpoint the individual variations from standard anatomical models. Methods We propose an automated procedure to create mechanical models of the human liver with patient-specific geometry and real time capabilities. The method hinges on the use of Statistical Shape Analysis to extract the relevant anatomical features from a database of medical images and Model Order Reduction to compute an explicit parametric solution for the mechanical response as a function of such features. The Sparse Subspace Learning, coupled with a Finite Element solver, was chosen to create low-rank solutions using a non-intrusive sparse sampling of the feature space. Results In the application presented in the paper, the statistical shape model was trained on a database of 385 three dimensional liver shapes, extracted from medical images, in order to create a parametrized representation of the liver anatomy. This parametrization and an additional parameter describing the breathing motion in linear elasticity were then used as input in the reduced order model. Results show a consistent agreement with the high fidelity Finite Element models built from liver images that were excluded from the training dataset. However, we evidence in the discussion the difficulty of having compact shape parametrizations arising from the extreme variability of the shapes found in the dataset and we propose potential strategies to tackle this issue. Conclusions A method to represent patient-specific real-time liver deformations during breathing is proposed in linear elasticity. Since the proposed method does not require any adaptation to the direct Finite Element solver used in the training phase, the procedure can be easily extended to more complex non-linear constitutive behaviors - such as hyperelasticity - and more general load cases. Therefore it can be integrated with little intrusiveness to generic simulation software including more sophisticated and realistic models. [ABSTRACT FROM AUTHOR]

Primary human hepatocytes (PHHs) remain the gold standard for in vitro investigations of xenobiotic metabolism and hepatotoxicity. However, scarcity of liver tissue and novel developments in liver surgery has limited the availability and quality of tissue samples. In particular, warm ischemia shifts the intracellular metabolism from aerobic to anaerobic conditions, which increases glycogenolysis, glucose depletion and energy deficiency. Therefore, the aim of the present study was to investigate whether supplementation with glucose and insulin during PHH isolation could reconstitute intracellular glycogen storage and beneficially affect viability and functionality. Furthermore, the study elucidated whether the susceptibility of the tissue's energy status correlates with body mass index (BMI). PHHs from 12 donors were isolated from human liver tissue obtained from partial liver resections using a two-step EDTA/collagenase perfusion technique. For a direct comparison of the influence of glucose/insulin supplementation, we modified the setup, enabling the parallel isolation of two pieces of one tissue sample with varying perfusate. Independent of the BMI of the patient, the glycogen content in liver tissue was notably low in the majority of samples. Furthermore, supplementation with glucose and insulin had no beneficial effect on the glycogen concentration of isolated PHHs. However, an indirect improvement of the availability of energy was shown by increased viability, plating efficiency and partial cellular activity after supplementation. The plating efficiency showed a striking inverse correlation with increasing lipid content of PHHs. However, 60 h of cultivation time revealed no significant impact on the maintenance of albumin and urea synthesis or xenobiotic metabolism after supplementation. In conclusion, surgical procedures and tissue handling may decrease hepatic energy resources and lead to cell stress and death. Consequently, PHHs with low energy resources die during the isolation process without supplementation of glucose/insulin or early cell culture, while their survival rates are improved with glucose/insulin supplementation. [ABSTRACT FROM AUTHOR]

Since the discovery of TRP proteins in 1969, during studies of the fruit fly Drosophila melanogaster, interest around them and the subfamily of TRPC channels has remained high. TRPC3 was able to be detected in a number of organs in rodents, such as rats and mice, and also in various human tissues. For the most part, these investigations were carried out using gene expression of TRPC3. Further work has already confirmed the relevance of TRPC3 in the context of neurodegenerative diseases, such as spinocerebellar ataxia, and carcinogenic entities, such as ovarian carcinoma. An association with TRPC3 has also been demonstrated for diseases that affect the liver. In order to confirm the expression of TRPC3 in the human liver, this study uses samples taken from eight (n = 8) fixated human body donors and analyzed with immunohistochemistry. In accordance with the macroscopic anatomy of the organs, six samples (n = 6) of liver tissue and three (n = 3) of gallbladder tissue were obtained. TRPC3 was clearly detected in all liver and gallbladder samples examined. Thus, it is not unlikely that TRPC3 plays a role in the extensive metabolic processes of the liver and could also serve as a target for pharmacological interventions in an imbalance of calcium homeostasis. [ABSTRACT FROM AUTHOR]

Liver injury is one of the most severe traffic trauma. The development of accurate liver finite element model is beneficial for improving the biofidelity and validity in crash simulations, aiming to analyze the injury in accidents. 12 human liver finite element models in the current research were constructed from high resolution CT data of a Chinese male 50th percentile human subject, including the main structures like left lobe, right lobe, capsule, parenchyma and falciform ligament. The simulations based on Nava et al. experiment were conducted to validate the models and make comparisons of modeling method accuracy. The results demonstrated that the larger deviation happened to the tetra models due to the stiffer algorithm compared to the hex models, which were more sensitive to element size. The existence of capsule had significant effects on the liver mechanical responses, reducing the liver tissue pressure. Shell elements were more suitable for modeling the capsule.

Background & Aims γδ T cells comprise a substantial proportion of tissue-associated lymphocytes. However, our current understanding of human γδ T cells is primarily based on peripheral blood subsets, while the immunobiology of tissue-associated subsets remains largely unclear. Therefore, we aimed to elucidate the T cell receptor (TCR) diversity, immunophenotype and function of γδ T cells in the human liver. Methods We characterised the TCR repertoire, immunophenotype and function of human liver infiltrating γδ T cells, by TCR sequencing analysis, flow cytometry, in situ hybridisation and immunohistochemistry. We focussed on the predominant tissue-associated Vδ2 − γδ subset, which is implicated in liver immunopathology. Results Intrahepatic Vδ2 − γδ T cells were highly clonally focussed, with single expanded clonotypes featuring complex, private TCR rearrangements frequently dominating the compartment. Such T cells were predominantly CD27 lo/− effector lymphocytes, whereas naïve CD27 hi , TCR-diverse populations present in matched blood were generally absent in the liver. Furthermore, while a CD45RA hi Vδ2 − γδ effector subset present in both liver and peripheral blood contained overlapping TCR clonotypes, the liver Vδ2 − γδ T cell pool also included a phenotypically distinct CD45RA lo effector compartment that was enriched for expression of the tissue tropism marker CD69, the hepatic homing chemokine receptors CXCR3 and CXCR6, and liver-restricted TCR clonotypes, suggestive of intrahepatic tissue residency. Liver infiltrating Vδ2 − γδ cells were capable of polyfunctional cytokine secretion, and unlike peripheral blood subsets, were responsive to both TCR and innate stimuli. Conclusion These findings suggest that the ability of Vδ2 − γδ T cells to undergo clonotypic expansion and differentiation is crucial in permitting access to solid tissues, such as the liver, which results in functionally distinct peripheral and liver-resident memory γδ T cell subsets. They also highlight the inherent functional plasticity within the Vδ2 − γδ T cell compartment and provide information that could be used for the design of cellular therapies that suppress liver inflammation or combat liver cancer. Lay summary γδ T cells are frequently enriched in many solid tissues, however the immunobiology of such tissue-associated subsets in humans has remained unclear. We show that intrahepatic γδ T cells are enriched for clonally expanded effector T cells, whereas naïve γδ T cells are largely excluded. Moreover, whereas a distinct proportion of circulating T cell clonotypes was present in both the liver tissue and peripheral blood, a functionally and clonotypically distinct population of liver-resident γδ T cells was also evident. Our findings suggest that factors triggering γδ T cell clonal selection and differentiation, such as infection, can drive enrichment of γδ T cells into liver tissue, allowing the development of functionally distinct tissue-restricted memory populations specialised in local hepatic immunosurveillance. [ABSTRACT FROM AUTHOR]

Abstract: Innate lymphocytes are selectively enriched in the liver where they have important roles in liver immunology. Murine studies have shown that type I NKT cells can promote liver inflammation, whereas type II NKT cells have an anti‐inflammatory role. In humans, type II NKT cells were found to accumulate in the gut during inflammation and IL13Rα2 was proposed as a marker for these cells. In the human liver, less is known about type I and II NKT cells. Here, we studied the phenotype and function of human liver T cells expressing IL13Rα2. We found that IL13Rα2 was expressed by around 1% of liver‐resident memory T cells but not on circulating T cells. In support of their innate‐like T‐cell character, the IL13Rα2+ T cells had higher expression of promyelocytic leukaemia zinc finger (PLZF) compared to IL13Rα2− T cells and possessed the capacity to produce IL‐22. However, only a minority of human liver sulfatide‐reactive type II NKT cells expressed IL13Rα2. Collectively, these findings suggest that IL13Rα2 identifies tissue‐resident intrahepatic T cells with innate characteristics and the capacity to produce IL‐22. [ABSTRACT FROM AUTHOR]

Chukanovite has been identified as an oxidation product of carbon steel in a human liver through powder X-ray powder diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM). Steel oxidation products include siderite, chukanovite, vivianite, magnetite, goethite, 2-line ferrihydrite. Hydroxylapatite and amorphous silica surrounding the oxidized steel are precipitated from body fluid. The Rietveld refinement of the chukanovite structure shows it is as expected monoclinic, space group of P 2 1 / a , with a  = 12.402(2), b  = 9.414(1), c  = 3.216(1) Å, β  = 97.73(3)°. The standard Gibbs free energy of formation of chukanovite is estimated at −1167.2 ± 1.2 kJ mol −1 from the equilibrium between siderite and chukanovite. By using linear free energy relationship, the Gibbs free energy of formation for other isostructural phases of pokrovskite, parádsasvárite, and nullaginite are predicted. The results of this study could improve the mineralogical and geochemical properties of chukanovite which is an oxidation product of carbon steel relevant also in radioactive waste containers and oil/gas pipes. Furthermore, the combined method of experimental measurements and geochemical modeling could be a useful tool to study the formation of inorganic solid precipitates and bio-materials in biofluids, and is especially important in medical mineralogy field. [ABSTRACT FROM AUTHOR]

A fast and accurate fusion of intra-operative images with a pre-operative data is a key component of computer-aided interventions which aim at improving the outcomes of the intervention while reducing the patient’s discomfort. In this paper, we focus on the problematic of the intra-operative navigation during abdominal surgery, which requires an accurate registration of tissues undergoing large deformations. Such a scenario occurs in the case of partial hepatectomy: to facilitate the access to the pathology, e.g. a tumor located in the posterior part of the right lobe, the surgery is performed on a patient in lateral position. Due to the change in patient’s position, the resection plan based on the pre-operative CT scan acquired in the supine position must be updated to account for the deformations. We suppose that an imaging modality, such as the cone-beam CT, provides the information about the intra-operative shape of an organ, however, due to the reduced radiation dose and contrast, the actual locations of the internal structures necessary to update the planning are not available. To this end, we propose a method allowing for fast registration of the pre-operative data represented by a detailed 3D model of the liver and its internal structure and the actual configuration given by the organ surface extracted from the intra-operative image. The algorithm behind the method combines the iterative closest point technique with a biomechanical model based on a co-rotational formulation of linear elasticity which accounts for large deformations of the tissue. The performance, robustness and accuracy of the method is quantitatively assessed on a control semi-synthetic dataset with known ground truth and a real dataset composed of nine pairs of abdominal CT scans acquired in supine and flank positions. It is shown that the proposed surface-matching method is capable of reducing the target registration error evaluated of the internal structures of the organ from more than 40 mm to less then 10 mm. Moreover, the control data is used to demonstrate the compatibility of the method with intra-operative clinical scenario, while the real datasets are utilized to study the impact of parametrization on the accuracy of the method. The method is also compared to a state-of-the art intensity-based registration technique in terms of accuracy and performance. [ABSTRACT FROM AUTHOR]

Abstract: Introduction: Murine hepatic NK cells exhibit adaptive features, with liver‐specific adhesion molecules CXCR6 and CD49a acting as surface markers. Methods: We investigated human liver‐resident CXCR6+ and CD49a+ NK cells using RNA sequencing, flow cytometry, and functional analysis. We further assessed the role of cytokines in generating NK cells with these phenotypes from the peripheral blood. Results: Hepatic CD49a+ NK cells could be induced using cytokines and produce high quantities of IFNγ and TNFα, in contrast to hepatic CXCR6+ NK cells. RNA sequencing of liver‐resident CXCR6+ NK cells confirmed a tolerant immature phenotype with reduced expression of markers associated with maturity and cytotoxicity. Liver‐resident double‐positive CXCR6 + CD49a+ hepatic NK cells are immature but maintain high expression of Th1 cytokines as observed for single‐positive CD49a+ NK cells. We show that stimulation with activating cytokines can readily induce upregulation of both CD49a and CXCR6 on NK cells in the peripheral blood. In particular, IL‐12 and IL‐15 can generate CXCR6 + CD49a+ NK cells in vitro from NK cells isolated from the peripheral blood, with comparable phenotypic and functional features to liver‐resident CD49a+ NK cells, including enhanced IFNγ and NKG2C expression. Conclusion: IL‐12 and IL‐15 may be key for generating NK cells with a tissue‐homing phenotype and strong Th1 cytokine profile in the blood, and links peripheral activation of NK cells with tissue‐homing. These findings may have important therapeutic implications for immunotherapy of chronic liver disease. [ABSTRACT FROM AUTHOR]

Valproic acid (VPA) is a very potent anti-cancer and neuro-protective drug probably by its HDAC inhibiting properties, which may cause steatosis in the liver. The present study investigates the effect of repetitive VPA treatment of primary human hepatocytes (PHH) on whole genome gene expression-, DNA methylation-, and miRNA changes, using microarrays and integrated data analyses. PHH were exposed to a non-cytotoxic dose of VPA for 5 days daily which induced lipid accumulation. Part of the PHH was left untreated for 3 days for studying the persistence of ‘omics’ changes. VPA treatment appeared to inhibit the expression of the transcription factors HNF1A and ONECUT1 . HNF1A interacted with 41 differentially expressed genes of which 12 were also differentially methylated. None of the genes present in this network were regulated by a DE-miR. The subnetwork of ONECUT1 consisted of 44 differentially expressed genes of which 15 were differentially methylated, and 3 were regulated by a DE-miR. A number of genes in the networks are involved in fatty acid metabolism, and may contribute to the development of steatosis by increasing oxidative stress thereby causing mitochondrial dysfunction, and by shifting metabolism of VPA towards β-oxidation due to reduced glucuronidation. Part of the changes remained persistent after washing out of VPA, like PMAIP1 which is associated with cellular stress in liver of patients with NASH. The MMP2 gene showed the highest number of interactions with other persistently expressed genes, among which LCN2 which is a key modulator of lipid homeostasis. Furthermore, VPA modulated the expression and DNA methylation level of nuclear receptors and their target genes involved in the adverse outcome pathway of steatosis, thereby expanding our current knowledge of the pathway. In particular, VPA modulated PPARγ, and PPARα, AHR and CD36 on both the gene expression and the DNA methylation level, thereby inhibiting β-oxidation and increasing uptake of fatty acid into the hepatocytes, respectively. Overall, our integrative data analyses identified novel genes modulated by VPA, which provide more insight into the mechanisms of repeated dose toxicity of VPA, leading to steatosis. [ABSTRACT FROM AUTHOR]

Decellularized human livers are considered the perfect extracellular matrix (ECM) surrogate because both three-dimensional architecture and biological features of the hepatic microenvironment are thought to be preserved. However, donor human livers are in chronically short supply, both for transplantation or as decellularized scaffolds, and will become even scarcer as life expectancy increases. It is hence of interest to determine the structural and biochemical properties of human hepatic ECM to derive design criteria for engineering biomimetic scaffolds. The intention of this work was to obtain quantitative design specifications for fabricating scaffolds for hepatic tissue engineering using human livers as a template. To this end, hepatic samples from five patients scheduled for hepatic resection were decellularized using a protocol shown to reproducibly conserve matrix composition and microstructure in porcine livers. The decellularization outcome was evaluated through histological and quantitative image analyses to evaluate cell removal, protein, and glycosaminoglycan content per unit area. Applying the same decellularization protocol to human liver samples obtained from five different patients yielded five different outcomes. Only one liver out of five was completely decellularized, while the other four showed different levels of remaining cells and matrix. Moreover, protein and glycosaminoglycan content per unit area after decellularization were also found to be patient- (or donor-) dependent. This donor-to-donor variability of human livers thus precludes their use as templates for engineering a generic 'one-size fits all' ECM-mimic hepatic scaffold. [ABSTRACT FROM AUTHOR]

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 = 0.43, and r = 0.44, respectively). Univariate and multivariate statistical analysis identified significant associations of SULT1A1 CNVs with mRNA ( p = 1.7 × 10) and protein ( p = 4.4 × 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. [ABSTRACT FROM AUTHOR]