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X X Mi, J Yan, X D Zeng, J P Shi
Objective: To establish overfed zebrafish model for non-alcoholic steatohepatitis. Methods: The wild-type zebrafish was fed 3 times a day with normal diet. Body length, weight, and triglyceride levels were measured after 20 days of feeding. The changes in expression of genes associated with cholesterol metabolism, lipid metabolism, endoplasmic reticulum stress, and inflammation were detected by quantitative PCR. Liver tissue sections were stained with H&E. Statistical analyses between groups were compared using t-test...
July 20, 2018: Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology
Y P Zhou, R Wu, W Shen, H H Yu, S J Yu
Objective: To explore the effects of oleic acid and palmitic acid on lipid deposition and mTOR/S6K1/SREBP-1c pathways in HepG2 cells. Methods: The model of steatosis was established with induction of oleic acid and palmitic acid and was intervened by rapamycin. The changes in lipid droplets were observed after staining the cells with oil Red O. Intracellular triglyceride (TG) contents in cells were measured by TG kit. mTOR, S6K1, and SREBP-1c mRNA expression levels were detected using QRT-PCR. Western blot was used to determine protein expression levels of mTOR, S6K1 and SREBP-1c...
June 20, 2018: Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology
S M Li, Z L Gao, X T Lyu, J Jia, P Zhu, Y Chen, Y L Liu, Z X Zhang
Objective: To investigate the diagnostic value of whole liver CT perfusion imaging in the quantitative evaluation of hemodynamic changes before and after transcatheter arterial chemoembolization (TACE). Methods: Twenty-six patients with hepatocellular carcinoma underwent TACE therapies were recruited. Whole -liver computed tomographic perfusion imaging (CTPI) was performed 2~3 days before TACE and 1 month after TACE. We measured the following perfusion parameters: hepatic arterial perfusion (HAP), portal venous perfusion (PVP), total liver perfusion (TLP), hepatic arterial perfusion index (HAPI), and time-to-peak (TTP)...
June 20, 2018: Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology
Sherif Abbas, Nihal Simsek Ozek, Salih Emri, Deniz Koksal, Mete Severcan, Feride Severcan
This study was conducted to differentiate malignant pleural mesothelioma (MPM) from lung cancer (LC) and benign pleural effusion (BPE) from pleural fluids using the diagnostic power of Fourier transform-infrared spectroscopy with attenuated total reflectance mode coupled with chemometrics. Infrared spectra of MPM (n  =  24), LC (n  =  20), and BPE (n  =  25) were collected, and hierarchical cluster analysis (HCA) and principal component analysis (PCA) were applied to their spectra. HCA results indicated that MPM was differentiated from LC with 100% sensitivity and 100% specificity and from BPE, with 100% sensitivity and 88% specificity, which were also confirmed by PCA score plots...
October 2018: Journal of Biomedical Optics
Mohammad N Oskouie, Nazanin S Aghili Moghaddam, Alexandra E Butler, Parvin Zamani, Amirhossein Sahebkar
Curcumin, the bioactive pigment of turmeric which has polyphenolic-hydrophobic components, has been used for the treatment of a variety of diseases. However, due to its insignificant intestinal-liver metabolism, low stability, quick systemic elimination and its hydrophobic property with low solubility, curcumin has limited bioavailability. Exosomes are nanovesicles (30-100 nm) released from diverse cell types into extracellular and, ultimately, into bio-fluids in a tightly regulated manner. Exosomes are capable of transferring lipids, proteins, RNAs and DNAs, both with and without direct cell-to-cell contact...
October 14, 2018: Journal of Cellular Physiology
Vahideh Tarhriz, Mojgan Bandehpour, Siavoush Dastmalchi, Elaheh Ouladsahebmadarek, Habib Zarredar, Shirin Eyvazi
Ovarian cancer (OC) is the fifth leading cause of cancer-related death among women. The high mortality rate is due to lack of early symptoms, late diagnosis, limited treatment options, and also emerging of drug resistance. Todays, molecular markers have become promising in tumor-targeted therapy. Several molecular markers have been known in OC immunotherapy. Identification of the specific molecular markers with prognostic significance is interested. CD24 is a small sialoglycoprotein which is localized in lipid rafts through its glycosylphosphatidylinositol (GPI) anchor...
October 14, 2018: Journal of Cellular Physiology
Edoardo Elia, Fabrizio Montecucco, Piero Portincasa, Amirhossein Sahebkar, Hamid Mollazadeh, Federico Carbone
Although coronary thrombosis (CT) is integral to cardiovascular outcomes, the underlying pathophysiological mechanisms remain unclear. CT may occur in case of atherosclerotic plaque erosion/rupture, or even after stenting implantation. Platelets (PLT) activation is the keystone of atherothrombosis and depends on many dysregulated elements, including endothelial dysfunction, oxidized lipoproteins, and immune response. Besides the classical view of PLT as an effector of hemostatic response, a new repertoire of PLT activities is emerging...
October 14, 2018: Journal of Cellular Physiology
Bahman Yousefi, Majid Eslami, Abdolmajid Ghasemian, Parviz Kokhaei, Amir Salek Farrokhi, Narges Darabi
Mammalian intestine contains a large diversity of commensal microbiota, which is far more than the number of host cells. Probiotics play an insecure and protective role against the colonization of intestinal pathogenic microbes and increase mucosal integrity by stimulating epithelial cells. Probiotics have innate capabilities in many ways, including receptor antagonism, receptor expression, binding and expression of adapter proteins, expression of negative regulatory signal molecules, induction of microRNAs, endotoxin tolerance, and ultimately secretion of immunomodulatory proteins, lipids, and metabolites to modulate the immune system...
October 14, 2018: Journal of Cellular Physiology
Liang Huang, Hongcheng Lin, Liang Kang, Pinzhu Huang, Jun Huang, Jinlin Cai, Zhenyu Xian, Peixuan Zhu, Meijin Huang, Liping Wang, Cory J Xian, Jianping Wang, Jianghui Dong
Long noncoding RNAs (lncRNAs) play a critical role in the initiation and progression of colorectal cancer (CRC), but little is known about the function of lncRNAs in the colorectal liver metastasis (CLM). This study was designed to identify specific lncRNAs correlating to liver metastasis of CRC, and to further assess their clinical value. Seventeen patients with primary CRC lesions, adjacent normal mucosa, and synchronous liver metastases lesions were divided into discovery set (six patients) and test set (11 patients)...
October 14, 2018: Journal of Cellular Physiology
Hai-Ming Xu, Feng-Hua Sui, Mei-Hua Sun, Gong-Liang Guo
Recent studies have shown that circulating microRNAs (miRNA) play a critical role in diagnosing acute coronary syndrome (ACS). This study aims to investigate the effect of miR-224 on atherosclerotic plaques forming and vascular remodeling in ACS and its relationship with TGF-β/Smad pathway. Myocardial infarction (MI) rat model was established and lentivirus vector of miR-224 inhibitor was prepared for investigating the effect of downregulated miR-224 on the contents of nitric oxide (NO) and endothelin-1 (ET-1), blood lipid levels and inflammatory factor levels in serum as well as the TGF-β/Smad pathway...
October 14, 2018: Journal of Cellular Physiology
Renato Chaves Souto Branco, Rafael Ludemann Camargo, Thiago Martins Batista, Jean Franciesco Vettorazzi, Camila Lubaczeuski, Lucas Henrique Montes Bomfim, Leonardo Reis Silveira, Antônio Carlos Boschero, Cláudio Cesar Zoppi, Everardo Magalhães Carneiro
Nutrient malnutrition, during the early stages of development, may facilitate the onset of metabolic diseases later in life. However, the consequences of nutritional insults, such as a high-fat diet (HFD) after protein restriction, are still controversial. We assessed overall glucose homeostasis and molecular markers of mitochondrial function in the gastrocnemius muscle of protein-restricted mice fed an HFD until early adulthood. Male C57BL/6 mice were fed a control (14% protein-control diet) or a protein-restricted (6% protein-restricted diet) diet for 6 weeks...
October 14, 2018: Journal of Cellular Physiology
Erdem Karatekin
Neurotransmitter and hormone release involve calcium-triggered fusion of a cargo-loaded vesicle with the plasma membrane. The initial connection between the fusing membranes, called the fusion pore, can evolve in various ways, including rapid dilation to allow full cargo release, slow expansion, repeated opening-closing, and resealing. Pore dynamics determine the kinetics of cargo release and the mode of vesicle recycling, but how these processes are controlled are poorly understood. Previous reconstitutions could not monitor single pores, limiting mechanistic insight they could provide...
October 13, 2018: FEBS Letters
Sevan Mattie, Tom Kazmirchuk, Jeannie Mui, Hojatollah Vali, Christopher Leonard Brett
SNARE-mediated membrane fusion is required for membrane trafficking as well as organelle biogenesis and homeostasis. The membrane fusion reaction involves sequential formation of hemifusion intermediates, whereby lipid monolayers partially mix on route to complete bilayer merger. Studies of the Saccharomyces cerevisiae lysosomal vacuole have revealed many of the fundamental mechanisms that drive the membrane fusion process, as well as features unique to organelle fusion. However, until recently, it has not been amenable to electron microscopy methods that have been invaluable for studying hemifusion in other model systems...
2019: Methods in Molecular Biology
Pei Li, Yawei Dai, Markus Seeger, Yan-Wen Tan
Fӧrster-type resonance energy transfer (FRET) with fluorescence cross-correlation spectroscopy (FCCS) is a powerful combination for observing intramolecular conformational dynamics on the micro- to millisecond timescale. Owing to its sensitivity to various physical parameters, FRET-FCCS has also been used to detect the reagent effects on proteins dynamics. However, FRET-FCCS alone cannot acquire the exact measurements of rate constants. Moreover, this technique is highly model dependent and can be unreliable when determining too many parameters at once...
2019: Methods in Molecular Biology
Yachong Hu, Zhiqi Tian, Jiajie Diao
This chapter expounds the single vesicle-vesicle fluorescence resonance energy transfer (FRET) measurement to study the membrane fusion mediated by SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor) proteins. The formation of a four-α-helix bundle of SNARE proteins can drive two membranes to a close proximity for fusion. Through single-molecule FRET-based microscopy, the lipid-mixing process at the single-vesicle level can be tracked in real time. This reconstitution system is applicable to study the molecular mechanism of SNAREs during different membrane fusion stages, such as docking, hemifusion, and full fusion...
2019: Methods in Molecular Biology
Joji Mima
Membrane fusion mediated by SNARE (soluble N-ethylmaleimide-sensitive factor attachment protein receptor)-family proteins is an essential process for intracellular membrane trafficking in all eukaryotic cells, which delivers proteins and lipids to their appropriate subcellular membrane compartments such as organelles and plasma membrane. The molecular basis of SNARE-mediated membrane fusion has been revealed by studying fusion of reconstituted proteoliposomes bearing purified SNARE-family proteins and chemically defined lipid species...
2019: Methods in Molecular Biology
Youngsoo Jun
SNARE complexes assembled between fusing membranes (in trans) are the core machinery driving lipid bilayer merger. Thus, an assay monitoring the formation of these trans-SNARE complexes is essential for SNARE-mediated membrane fusion studies. Homotypic yeast vacuole fusion is an important model system for such studies. Although several assays measuring trans-SNARE complex formation are available to study yeast vacuole fusion, most use SNAREs conjugated with epitope tags, which may affect the function of SNAREs or even the formation of trans-SNARE complexes...
2019: Methods in Molecular Biology
Massimo D'Agostino, Andreas Mayer
Fluorescence de-quenching can be used to analyze membrane lipid mixing during an in vitro fusion reaction. Here we describe a method to measure lipid mixing using vacuolar membranes purified from the yeast Saccharomyces cerevisiae. Labeling the isolated organelles with rhodamine-phosphatidylethanolamine allows to reveal ATP-dependent lipid mixing through fluorescence de-quenching in a spectrofluorometer. Combining this assay with content mixing indicators, such as the fusion-dependent maturation of a luminal vacuolar phosphatase, then permits the detection of hemifusion intermediates and the analysis of the requirements for fusion pore opening...
2019: Methods in Molecular Biology
Matthew L Starr, Rutilio Fratti
Protein-lipid binding interactions play a key role in the regulation of peripheral membrane protein function. Liposome-binding assays are a simple and affordable means of screening for specific protein-lipid interactions. Liposomes are prepared by mixing phospholipid combinations of interest before drying and rehydration. Sonication of the lipid mixture produces small unilamellar vesicles (SUVs) which are incubated with a protein of interest to allow for any binding to occur. Liposomes and liposome-protein complexes are floated on a sucrose gradient by centrifugation to separate them from unbound protein...
2019: Methods in Molecular Biology
Robert P Sparks, Jermaine L Jenkins, Rutilio Fratti
Surface plasmon resonance (SPR) can be used to analyze both binding affinities and kinetic parameters between a ligand and an analyte. SPR can be performed by either cross-linking a given ligand to a sensor chip covalently or utilizing high-affinity non-covalent interactions to secure a ligand in a particular conformation to a chip, both of which have their potential advantages. SPR measurements are mass based and reflect the proportional amount of analyte bound to a given ligand at a given concentration when flowed at a set rate in order to determine the binding parameters of a given biochemical interaction...
2019: Methods in Molecular Biology
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