Subzero non-frozen preservation of human livers in the supercooled state

Using chicken antibodies IgY (purified from egg yolks) against mammalian cytochromes P450 and by means of cytochrome P450 marker substrates, we found for the first time the presence of hepatopancreatic cytochrome P450 in crayfishOrconectes limosus(an inducible cytochrome P450 2B-like enzyme) and we were able to detect and quantify cytochrome P450 1A1 in microsomes of human livers. Expression levels of cytochrome P450 1A1 in human livers constituted less than 0.6% of the total hepatic cytochrome P450 complement. The results obtained in our study are clear examples that chicken IgY are suitable for cytochrome P450 detection and quantification. Due to the evolutionary distance, chicken IgY reacts with more epitopes on a mammalian antigen, which gives an amplification of the signal. Moreover, this approach offers many advantages over common mammalian antibody production since chicken egg is an abundant source of antibodies (about 100 mg IgY/yolk) and the egg collection is a non-invasive technique. In the case of antibodies against cytochrome P450 2B4, we documented fast and steady production of highly specific immunoglobulins. Thus, chicken antibodies should be considered as a good alternative to and/or superior substitute for conventional polyclonal antibody produced in mammals.

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A beautiful day in the neighborhood: Application of single cell transcriptomics to unravel liver cell heterogeneity in diseased human livers

Hepatology ◽

10.1002/hep.31827 ◽

2021 ◽

Author(s):

Tianyi Chen ◽

Anna Mae Diehl

Keyword(s):

Single Cell ◽

Liver Cell ◽

Cell Heterogeneity ◽

Human Livers

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Expression of cytochrome P450 isozyme transcripts and activities in human livers

Xenobiotica ◽

10.1080/00498254.2020.1867929 ◽

2020 ◽

pp. 1-8

Author(s):

Jie Liu ◽

Yuan-Fu Lu ◽

J. Christopher Corton ◽

Curtis D. Klaassen

Keyword(s):

Cytochrome P450 ◽

Human Livers

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Variation in the expression of cytochrome P450-related miRNAs and transcriptional factors in human livers: Correlation with cytochrome P450 gene phenotypes

Toxicology and Applied Pharmacology ◽

10.1016/j.taap.2020.115389 ◽

2021 ◽

Vol 412 ◽

pp. 115389

Author(s):

Hai-feng Zhang ◽

Li-li Zhu ◽

Xiao-bei Yang ◽

Na Gao ◽

Yan Fang ◽

...

Keyword(s):

Cytochrome P450 ◽

Transcriptional Factors ◽

Cytochrome P450 Gene ◽

P450 Gene ◽

Human Livers

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CHOP and c-JUN up-regulate the mutant Z α1-antitrypsin, exacerbating its aggregation and liver proteotoxicity

Journal of Biological Chemistry ◽

10.1074/jbc.ra120.014307 ◽

2020 ◽

Vol 295 (38) ◽

pp. 13213-13223

Author(s):

Sergio Attanasio ◽

Rosa Ferriero ◽

Gwladys Gernoux ◽

Rossella De Cegli ◽

Annamaria Carissimo ◽

...

Keyword(s):

Liver Disease ◽

Er Stress ◽

Proteinase Inhibitors ◽

Pediatric Population ◽

Transcriptional Response ◽

Luciferase Reporter ◽

Mrna Levels ◽

Primary Role ◽

Loss Of Function ◽

Human Livers

α1-Antitrypsin (AAT) encoded by the SERPINA1 gene is an acute-phase protein synthesized in the liver and secreted into the circulation. Its primary role is to protect lung tissue by inhibiting neutrophil elastase. The Z allele of SERPINA1 encodes a mutant AAT, named ATZ, that changes the protein structure and leads to its misfolding and polymerization, which cause endoplasmic reticulum (ER) stress and liver disease through a gain-of-function toxic mechanism. Hepatic retention of ATZ results in deficiency of one of the most important circulating proteinase inhibitors and predisposes to early-onset emphysema through a loss-of-function mechanism. The pathogenetic mechanisms underlying the liver disease are not completely understood. C/EBP-homologous protein (CHOP), a transcription factor induced by ER stress, was found among the most up-regulated genes in livers of PiZ mice that express ATZ and in human livers of patients homozygous for the Z allele. Compared with controls, juvenile PiZ/Chop−/− mice showed reduced hepatic ATZ and a transcriptional response indicative of decreased ER stress by RNA-Seq analysis. Livers of PiZ/Chop−/− mice also showed reduced SERPINA1 mRNA levels. By chromatin immunoprecipitations and luciferase reporter–based transfection assays, CHOP was found to up-regulate SERPINA1 cooperating with c-JUN, which was previously shown to up-regulate SERPINA1, thus aggravating hepatic accumulation of ATZ. Increased CHOP levels were detected in diseased livers of children homozygous for the Z allele. In summary, CHOP and c-JUN up-regulate SERPINA1 transcription and play an important role in hepatic disease by increasing the burden of proteotoxic ATZ, particularly in the pediatric population.

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Plasmodium falciparum infection and exoerythrocytic development in mice with chimeric human livers

International Journal for Parasitology ◽

10.1016/j.ijpara.2005.10.014 ◽

2006 ◽

Vol 36 (3) ◽

pp. 353-360 ◽

Cited By ~ 73

Author(s):

John B. Sacci ◽

Uzma Alam ◽

Donna Douglas ◽

Jamie Lewis ◽

D Lorne J. Tyrrell ◽

...

Keyword(s):

Plasmodium Falciparum ◽

Falciparum Infection ◽

Plasmodium Falciparum Infection ◽

Human Livers

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Quantitation of replication of the HCV genome in human livers with end-stage cirrhosis by strand-specific real-time RT-PCR assays: Methods and clinical relevance

Journal of Medical Virology ◽

10.1002/jmv.21510 ◽

2009 ◽

Vol 81 (9) ◽

pp. 1569-1575 ◽

Cited By ~ 4

Author(s):

Lan Lin ◽

Louis Libbrecht ◽

Jannick Verbeeck ◽

Chris Verslype ◽

Tania Roskams ◽

...

Keyword(s):

Real Time ◽

Clinical Relevance ◽

Rt Pcr ◽

Pcr Assays ◽

Human Livers ◽

End Stage

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Energy determinants GAPDH and NDPK act as genetic modifiers for hepatocyte inclusion formation

The Journal of Cell Biology ◽

10.1083/jcb.201102142 ◽

2011 ◽

Vol 195 (2) ◽

pp. 217-229 ◽

Cited By ~ 23

Author(s):

Natasha T. Snider ◽

Sujith V.W. Weerasinghe ◽

Amika Singla ◽

Jessica M. Leonard ◽

Shinichiro Hanada ◽

...

Keyword(s):

Liver Injury ◽

Human Liver ◽

Nuclear Translocation ◽

Nucleoside Diphosphate Kinase ◽

Isolated Hepatocytes ◽

Dependent Manner ◽

Genetic Modifiers ◽

C3h Mice ◽

Normal Human ◽

Human Livers

Genetic factors impact liver injury susceptibility and disease progression. Prominent histological features of some chronic human liver diseases are hepatocyte ballooning and Mallory-Denk bodies. In mice, these features are induced by 3,5-diethoxycarbonyl-1,4-dihydrocollidine (DDC) in a strain-dependent manner, with the C57BL and C3H strains showing high and low susceptibility, respectively. To identify modifiers of DDC-induced liver injury, we compared C57BL and C3H mice using proteomic, biochemical, and cell biological tools. DDC elevated reactive oxygen species (ROS) and oxidative stress enzymes preferentially in C57BL livers and isolated hepatocytes. C57BL livers and hepatocytes also manifested significant down-regulation, aggregation, and nuclear translocation of glyceraldehyde 3-phosphate dehydrogenase (GAPDH). GAPDH knockdown depleted bioenergetic and antioxidant enzymes and elevated hepatocyte ROS, whereas GAPDH overexpression decreased hepatocyte ROS. On the other hand, C3H livers had higher expression and activity of the energy-generating nucleoside-diphosphate kinase (NDPK), and knockdown of hepatocyte NDPK augmented DDC-induced ROS formation. Consistent with these findings, cirrhotic, but not normal, human livers contained GAPDH aggregates and NDPK complexes. We propose that GAPDH and NDPK are genetic modifiers of murine DDC-induced liver injury and potentially human liver disease.

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