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Hepatocyte Isolation

Ekaterina S Kharechkina, Anna B Nikiforova, Vera V Teplova, Irina V Odinokova, Olga V Krestinina, Yulia L Baburina, Svetlana A Kruglova, Alexey G Kruglov
BACKGROUND: The opening of the permeability transition pore (PTP) in mitochondria plays a critical role in the pathogenesis of numerous diseases. Mitochondrial matrix pyridine nucleotides are potent regulators of the PTP, but the role of extramitochondrial nucleotides is unclear. METHODS: The PTP opening was explored in isolated mitochondria and mitochondria in permeabilized differentiated and undifferentiated cells in the presence of added NAD(P)(H) in combination with Mg2+ , adenine nucleotides (AN), and the inhibitors of AN translocase (ANT), voltage-dependent anion channel (VDAC), and cyclophilin D...
February 11, 2019: Biochimica et biophysica acta. General subjects
Ramla Sana Khalid, Irfan Khan, Midhat Batool Zaidi, Nadia Naeem, Kanwal Haneef, Rida-E-Maria Qazi, Rakhshinda Habib, Tuba Shakil Malick, Anwar Ali, Asmat Salim
This study was aimed to enhance the healing potential of rat bone marrow mesenchymal stem cells against chronic diabetic wounds through interleukin-7 (IL-7) transfection. IL-7 plays an important role in wound healing and acts as a survival factor in some cell types. This study involves isolation, propagation and characterization of MSCs and their modification with IL-7 gene via retroviral transfection. Transfected MSCs were assessed for their effect on angiogenic genes by qPCR. Wound healing potential of transfected MSCs was analyzed by scratch assay in vitro and by transplanting these cells in rat diabetic wound models in vivo...
February 13, 2019: Wound Repair and Regeneration
Tyler D Bammert, Collin A Beckstrom, Grace Lincenberg, Jamie G Hijmans, Jared J Greiner, Natalia G Rocha
Bone marrow-derived early outgrowth cells play an important role in endothelial repair. In vitro isolation techniques have identified two distinct morphological early outgrowth cell populations, but it is still unknown whether they present some functional phenotypic differences. Accordingly, the aim of the present study was to determine whether there are phenotypic differences in cellular function between two putative early outgrowth cells in culture. Peripheral blood samples were collected from 18 healthy adults...
February 12, 2019: Cytotechnology
Kelly McDaniel, Nan Wu, Tianhao Zhou, Li Huang, Keisaku Sato, Julie Venter, Ludovica Ceci, Demeng Chen, Sugeily Ramos-Lorenzo, Pietro Invernizzi, Francesca Bernuzzi, Chaodong Wu, Heather Francis, Shannon Glaser, Gianfranco Alpini, Fanyin Meng
Cholangiopathies are diseases that affect cholangiocytes, the cells lining the biliary tract. Liver stem cells (LSCs) are able to differentiate into all cells of the liver and possibly influence the surrounding liver tissue by secretion of signaling molecules. One way in which cells can interact is through secretion of extracellular vesicles (EVs), which are small membrane-bound vesicles, containing proteins, miRNAs and cytokines. We evaluated contents of liver stem cell derived EVs (LSCEVs), compared their miRNA contents to those of EVs isolated from hepatocytes, and evaluated the downstream targets of these miRNAs...
February 5, 2019: Hepatology: Official Journal of the American Association for the Study of Liver Diseases
Eric Cassmann, Bianca Zaffarano, Qi Chen, Ganwu Li, Joseph Haynes
A 15-year-old female cockatiel (Nymphicus hollandicus) undergoing long term management for hepatopathy died and underwent necropsy. Microscopic findings were consistent with chronic liver disease characterized by distorted hepatic architecture, fibrosis and biliary proliferation. The additional finding of large intranuclear inclusion bodies within hepatocytes and renal tubular epithelium prompted diagnostic next generation sequencing. The assembled sequences isolated from pooled kidney and liver were related to siadenoviruses...
January 31, 2019: Virus Research
Jana C Mossanen, Marlene Kohlhepp, Alexander Wehr, Oliver Krenkel, Anke Liepelt, Anjali A Roeth, Diana Möckel, Felix Heymann, Twan Lammers, Nikolaus Gassler, Juliane Hermann, Joachim Jankowski, Ulf P Neumann, Tom Luedde, Christian Trautwein, Frank Tacke
BACKGROUND & AIMS: Inflammation in the liver provokes fibrosis but inflammation is also important for tumor surveillance. Inhibitors of chemokine pathways, such as CXCL16 and CXCR6 regulation of lymphocyte trafficking, are being tested as anti-fibrotic agents, but their effects on development of hepatocellular carcinoma (HCC) are unclear. We assessed the roles of CXCR6-dependent immune mechanisms in hepatocarcinogenesis. METHODS: C57BL/6J wild-type (WT) mice and CXCR6-deficient mice (Cxcr6eGfp/eGfp ) were given injections of diethylnitrosamine (DEN) to induce liver cancer, and α-galactosylceramide to activate natural killer T (NKT) cells...
January 30, 2019: Gastroenterology
Antonio Gnoni, Luisa Siculella, Giuseppina Paglialonga, Fabrizio Damiano, Anna Maria Giudetti
Hepatic de novo lipogenesis (DNL), the process by which carbohydrates are converted into lipids, is strictly controlled by nutritional and hormonal status. 3,5-Diiodo-L-thyronine (T2), a product of the 3,5,3'-triiodo-L-thyronine (T3) peripheral metabolism, has been shown to mimic some T3 effects on lipid metabolism by a short-term mechanism independent of protein synthesis. Here, we report that T2, administered for 1 week to hypothyroid rats, increases total fatty acid synthesis from acetate in isolated hepatocytes...
February 1, 2019: IUBMB Life
Karuna Rasineni, Paul G Thomes, Jacy L Kubik, Edward Noel Harris, Kusum K Kharbanda, Carol A Casey
Fatty liver is the earliest response of the liver to excessive ethanol consumption. Central in the development of alcoholic steatosis is increased mobilization of non-esterified free fatty acids (NEFA) to the liver from the adipose tissue. In this study, we hypothesized that ethanol-induced increase in ghrelin by impairing insulin secretion, could be responsible for the altered lipid metabolism observed in adipose and liver tissue. Male Wistar rats were fed for 5 to 8 weeks with control or ethanol Lieber-DeCarli diet, followed by biochemical analyses in serum and liver tissues...
January 31, 2019: American Journal of Physiology. Gastrointestinal and Liver Physiology
Niu-Niu Zhang, Yun-Xiang Tang, Lu Qian, Ya-Min Gao, Zhi-Yong Liu, Zhi-Liang Zou, Tian-Yu Zhang, Ming Yan
A series of novel 3-amidophenols with 5-heteroatomic substitutions were designed and synthesized. Several compounds showed potent antitubercular activity against Mycobacterium tuberculosis H37Ra (MIC = 0.25-5 μg/mL). Compounds 12j and 14i also displayed good inhibitory activity against M. tuberculosis H37Rv and two clinically isolated multidrug-resistant M. tuberculosis strains (MIC = 0.39-3.12 μg/mL). The privileged compound 14i showed certain oral efficacy on a mouse infection model. The compounds are non-cytotoxic against L-O2 hepatocytes and RAW264...
January 30, 2019: Archiv der Pharmazie
Jessica Mae Grindheim, Dario Nicetto, Greg Donahue, Kenneth S Zaret
BACKGROUND & AIMS: Little is known about mechanisms of postnatal hepatocyte maturation or chromatin regulation of genes that control fibrogenesis. We investigated transcription of genes that regulate fibrosis and the effects of chromatin compaction and the polycomb repressive complex 2 (PRC2) in postnatal hepatocytes of mice, focusing on the roles of the histone methyltransferases EZH1 and EZH2. METHODS: Hepatocytes were isolated from C57BL/6J and C3H mice, as well as mice with liver-specific disruption of Ezh1 and/or Ezh2, at postnatal day 14 (P14) and 2 months after birth (M2)...
January 25, 2019: Gastroenterology
Alicia Maclaine, María J Forzán, Narges Mashkour, Jennifer Scott, Ellen Ariel
Juvenile eastern water dragons ( Intellagama lesueurii lesueurii) are highly susceptible to infection with Bohle iridovirus (BIV), a species of ranavirus first isolated from ornate burrowing frogs in Townsville, Australia. To investigate the progression of BIV infection in eastern water dragons, 11 captive-bred juveniles were orally inoculated with a dose of 104.33 TCID50 and euthanized at 3, 6, 8, 10, 12, and 14 days postinfection (dpi). Viral DNA was detected via polymerase chain reaction (PCR) in the liver, kidney, and cloacal swabs at 3 dpi...
January 27, 2019: Veterinary Pathology
Jiaxiang Zhang, Na Li, Ling Yang, Haibo Xie, Yi Yang, Hui Wang, Changhao Wu, Tong Shen, Qixing Zhu
We have previously shown trichloroethylene (TCE) induced occupational medicamentosa-like dermatitis due to TCE (OMLDT) with immune liver injury, and kallikrein-kinin system (KKS) activation as a probably mechanism underlying the immune damage. Bradykinin (BK) is an important active component of KKS system function, but the specific role of BK in the immune liver injury has never been examined. The present study aimed to explore the important role of BK and mechanisms of action in immune liver injury induced by TCE...
January 24, 2019: Toxicology
Guillaume Lassailly, Mohamed Bou Saleh, Natascha Leleu-Chavain, Massih Ningarhari, Emilie Gantier, Rodolphe Carpentier, Florent Artru, Viviane Gnemmi, Benjamin Bertin, Patrice Maboudou, Didier Betbeder, Céline Gheeraert, François Maggiotto, Sébastien Dharancy, Philippe Mathurin, Alexandre Louvet, Laurent Dubuquoy
OBJECTIVE: In liver transplantation, organ shortage leads to use marginal grafts that are more susceptible to ischemia reperfusion (IR). We identified nucleotide-binding oligomerization domain 1 (NOD1) as an important modulator of polymorphonuclear (PMN)-induced liver injury such as in IR. Here we aimed at elucidating its role in the liver IR, especially on the endothelium and hepatocyte. DESIGN: NOD1 WT and KO mice were treated with NOD1 agonists and submitted to liver IR...
January 24, 2019: Journal of Hepatology
Chunxia Gao, Fengzhong Wang, Li Yuan, Junyi Liu, Dan Sun, Xuyan Li
Appropriate germination can improve the nutritional value and bioactivity of soybeans; however, few studies have assessed the effect of germination on soybean proteins. This study examined the physicochemical property, antioxidation, and cytoprotective effect of the germinated soybean proteins (Gsp). Gsp was extracted from soybeans which germinated for 0-3 days using the method of alkali-solution and acid-isolation extraction. The results showed that germination could digest soybean proteins into the smaller molecules; enhance the degree of hydrolysis, emulsifiability, and foaming capacity; increase the removal rate of ABTS, DPPH, <mml:math xmlns:mml="http://www...
January 2019: Food Science & Nutrition
Maisa Alnaqeeb, Kenza A Mansor, Eyad M Mallah, Bayan Y Ghanim, Nasir Idkaidek, Nidal A Qinna
BACKGROUND: In-depth information of potential drug-herb interactions between warfarin and herbal compounds with suspected anticoagulant blood thinning effects is needed to raise caution of concomitant administration. The current study aimed to investigate the impact of co-administration of pomegranate peel and guava leaves extracts, including their quality markers namely; ellagic acid and quercetin, respectively, on warfarin's in vivo dynamic activity and pharmacokinetic actions, in addition to potential in vitro cytochrome P450 enzymes (CYP) inhibition...
January 24, 2019: BMC Complementary and Alternative Medicine
Qingyu Yang, Qi Zhang, Xuewu Zhang, Lei You, Wenbiao Wang, Weiyong Liu, Yang Han, Chunqiang Ma, Wei Xu, Junbo Chen, Hua Yang, Pin Wan, Yao Zhou, Yingle Liu, Kailang Wu, Ziwen Yang, Jianguo Wu
Hepatitis B virus (HBV) infection is the leading cause of chronic hepatitis B (CHB), liver cirrhosis (LC), and hepatocellular carcinoma (HCC). This study reveals a distinct mechanism underlying the regulation of HBV replication. HBV activates the homeobox A10 (HoxA10) in human hepatocytes, leukocytes, peripheral blood mononuclear cells (PBMCs), HepG2-NTCP cells, leukocytes isolated from CHB patients, and HBV-associated HCC tissues. HoxA10 in turn represses HBV replication in human hepatocytes, HepG2-NTCP cells, and BABL/c mice...
January 23, 2019: Journal of Virology
Xu Wang, Xin Xin, Ying Sun, Lizhu Zou, Hui Li, Yufei Zhao, Ruihong Li, Ying Peng, Jiang Zheng
Aloe-emodin (AE), an anthraquinone derivative, is a bioactive ingredient isolated from Rhubarb which is used to treat inflammatory illnesses in China and many other countries in Asia. AE has shown a wide range of pharmacological effects. Recent studies showed that exposure to AE could cause DNA damage and cytotoxicity. The goals of the present study aimed at 1) exploration of oxidative metabolism pathways of AE, 2) identification of P450 enzymes which respond the hydroxylation of AE, and 3) determination of electrophilicity of AE and its oxidative metabolites...
January 23, 2019: Chemical Research in Toxicology
Naoki Tanaka, Takefumi Kimura, Naoyuki Fujimori, Tadanobu Nagaya, Michiharu Komatsu, Eiji Tanaka
Non-alcoholic fatty liver disease (NAFLD) is a major chronic liver disease that can lead to liver cirrhosis, liver cancer, and ultimately death. NAFLD is pathologically classified as non-alcoholic fatty liver (NAFL) or non-alcoholic steatohepatitis (NASH) based on the existence of ballooned hepatocytes, although the states have been known to transform into each other. Moreover, since the detection of ballooned hepatocytes may be difficult with limited biopsied specimens, its clinical significance needs reconsideration...
January 14, 2019: World Journal of Gastroenterology: WJG
Weiyan Zhang, Yue Sun, Wei Liu, Jinling Dong, Jinglong Chen
Non‑alcoholic fatty liver disease (NAFLD) is the consequence of insulin resistance, fatty acid accumulation, oxidative stress and lipotoxicity. The present study aimed to elucidate the effect of Quaking 5 (QKI 5) as mediated by Sirtuin 1 (SIRT1) on triglyceride (TG) synthesis in the liver of an NAFLD mouse model. A high‑fat diet‑induced NAFLD model was established in mice, and mouse hepatocytes were isolated to characterize the effects of QKI 5 mediated by SIRT1 on TG synthesis in the liver. Body weight and liver wet weight were recorded...
January 10, 2019: International Journal of Molecular Medicine
Yan-Fang Tan, Lian Tang, Wen-Xian OuYang, Tao Jiang, Hui Zhang, Shuang-Jie Li
OBJECTIVE: To investigate role of β-catenin and lncRNA MALAT1/miR-217 axis to converge into the regulation of ZEB-1 in hepatocyte growth factor (HGF)-induced hepatocytes differentiated from bone marrow mesenchymal stem cells (BM-MSCs). METHODS: BM-MSCs were isolated and HGF was used to induce the differentiation of BM-MSCs into hepatocytes. HSC-T6 cells, BRL-3 A cells and differentiated BM-MSCs were treated by lipopolysaccharide(LPS). shRNAs were used to silence β-catenin and recombinant plasmids were used to over-express ZEB1...
January 7, 2019: Pathology, Research and Practice
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