bluuebunny/crossref_metadata_2025_split huggingface dataset:
Modulation of CYP3A4 activity and induction of apoptosis, necrosis and senescence by the anti‐tumour imidazoacridinone C‐1311 in human hepatoma cells
Authors: Augustin Ewa , Pawłowska Monika , Polewska Joanna , Potega Agnieszka , Mazerska Zofia 
Published: August 2013

AbstractThere is increasing evidence that the expression level of drug metabolic enzymes affects the final cellular response following drug treatment. Moreover, anti‐tumour agents may modulate enzymatic activity and/or cellular expression of metabolic enzymes in tumour cells. We have investigated the influence of CYP3A4 overexpression on the cellular response induced by the anti‐tumour agent C‐1311 in hepatoma cells. C‐1311‐mediated CYP3A4 activity modulation and the effect of CYP3A4 overexpression on C‐1311 metabolism have also been examined. With the HepG2 cell line and its CYP3A4‐overexpressing variant, Hep3A4, experiments involving DAPI staining, cell cycle analysis, phosphatidylserine externalisation and senescence‐associated (SA)‐β‐galactosidase expression, were used to monitor the effects of C‐1311 exposure. C‐1311 cellular metabolism and CYP3A4 activity were investigated by high‐performance liquid chromatography. C‐1311 metabolism was very low in both hepatoma cell lines and slightly influenced by CYP3A4 expression. Interestingly, in HepG2 cells, C‐1311 was an effective modulator of CYP3A4 enzymatic activity, being the inhibitor of this isoenzyme in Hep3A4 cells. Cell cycle analysis showed that HepG2 cells underwent a rather stable G2/M arrest following C‐1311 exposure, whereas CYP3A4‐overexpressing cells accumulated only slightly in this compartment. C‐1311‐treated cells died by apoptosis and necrosis, whereas surviving cells underwent senescence; however, these effects occurred faster and more intensely in Hep3A4 cells. Although CYP3A4 did not influence C‐1311 metabolism, changes in CYP3A4 levels affected the C‐1311‐induced response in hepatoma cells. Therefore, inter‐patient differences in CYP3A4 levels should be considered when assessing the potential therapeutic effects of C‐1311.
Isolation and in vitro propagation of human skeletal muscle progenitor cells from fetal muscle
Authors: Hosoyama Tohru , Meyer Michael G. , Krakora Dan , Suzuki Masatoshi 
Published: August 2013

AbstractSkeletal muscle progenitor cells (SMPCs) are considered one of the most valuable cells for cell‐based therapy targeting skeletal muscle. However, an efficient protocol for isolating and maintaining human myogenic progenitors in vitro has not been fully established. In this study, we demonstrate that human myogenic progenitors can be expanded and proliferated from human fetal muscles. Human SMPCs were prepared from fetal hind limb muscles and induced to proliferate as free‐floating spheres termed myospheres in the medium containing basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF). Both myogenic progenitors and myoblast populations from human fetal muscles were effectively propagated in myospheres and passaged by a mechanical chopping. After expanding these spheres in culture, we tested whether myogenic progenitor cells can differentiate into multinucleated myotubes. The myospheres were dissociated, plated down on coverslips and cultured in the medium for terminal differentiation. We could confirm that the plated cells formed well‐developed, multinucleated myotubes. This culture method using myospheres is an effective protocol to isolate and maintain SMPCs from human fetal skeletal muscles in culture.
Pharmacological doses of melatonin induce alterations in mitochondrial mass and potential, bcl‐2 levels and K+ currents in UVB‐exposed U937 cells
Authors: Canonico Barbara , Luchetti Francesca , Ambrogini Patrizia , Arcangeletti Marcella , Betti Michele , Cesarini Erica , Lattanzi Davide , Ciuffoli Stefano , Palma Fulvio , Cuppini Riccardo , Papa Stefano 
Published: August 2013

AbstractApoptosis is observed in ‘actively’ dying cells after the exposure to cell stressors such as ultraviolet light irradiation. Since melatonin has been proposed to act under stressful conditions as cell protection factor, in this study we examined the potential of this molecule when used at pharmacological concentrations to control mitochondrial damage and apoptotic signalling of UVB irradiated U937 human leukaemic cells. Moreover, the effect of melatonin treatment on electrophysiological properties and membrane K+ currents of irradiated U937 cells was investigated as functional aspects relevant to the anti‐apoptotic role of melatonin. The general effect is associated with the restoration of mass, number and membrane potential of mitochondria, with a lower caspase activation and bcl‐2 upregulation. In the presence of the caspase inhibitor ZVAD‐Fmk, melatonin seems to drive UVB stressed cells to follow the mitochondrial intrinsic pathway, interfering just at the mitochondrial level. Moreover, treatment with melatonin, as well as ZVAD‐Fmk, prevented the K+ current reduction observed late following the UVB insult application, by sparing cells from death; this result also indicates that the decrease of K+ leakage currents could represent a functional feature of apoptotic process in UV‐exposed U937 cells.
Evidence of chondrocyte turnover in lung cartilage, with the probable participation of nestin‐positive cells
Authors: Ortega‐Martínez Marta , Romero‐Núñez Edgar , Niderhauser‐García Alberto , de‐la‐Garza‐González Carlos , Ancer‐Rodríguez Jesús , Jaramillo‐Rangel Gilberto 
Published: August 2013

AbstractHealthy adult cartilage is thought to have little or no capacity to renewal, and cell turnover has not been reported in lung cartilage. We report that chondrocyte turnover occurs in lung cartilage, found in an unrelated study. Lung specimens from CD1 mice of 2, 6, 12, 18 or 24 months were fixed in 10% neutral‐buffered formalin and paraffin‐embedded. Apoptosis was analysed by in situ end‐labelling of fragmented DNA. Proliferating cell nuclear antigen (PCNA) and nestin were examined by immunohistochemistry. Apoptosis and PCNA were detected in lung chondrocytes. Serial section analysis showed that cells in apoptosis were different from PCNA‐positive cells, indicating that turnover was occurring. Chondrocytes were negative for nestin. Nestin‐positive cells were present in connective tissue associated with cartilage, in some specimens in close proximity of it and in perivascular cells. Thus cell turnover in lung cartilage is possible, which may be mediated by nestin‐positive cells.
Comparison of human first and third trimester placental mesenchymal stem cell
Authors: Park Saeyoung , Koh Seong‐Eun , Hur Chang Young , Lee Won‐Don , Lim Jinho , Lee Young‐Jay 
Published: August 2013

AbstractPlacenta mesenchymal stem cells (PMSCs) have the characteristic features of stem cells including renewability in vitro, surface expression, differentiation potency and ability to adhere to the culture surface. PMSCs expressed genes are normally found in the embryonic tissues before the onset of gastrulation, indicating multipotency. However, the stemness can depend on the stages of the placenta from which the cells were isolated. PMSCs were isolated from two different stages of placenta for comparison, that is the first and third trimesters. Both sets had very similar patterns of surface expression as CD44, CD73, CD90 and CD105, and of self renewability in vitro. Expressions of pluripotency‐coupled genes were also confirmed in both sets of cells; however, there was a significant difference in the expression levels: fPMSC (mesenchymal stem cells isolated from the first trimester human placenta) being 2–11‐fold higher than tPMSC (mesenchymal stem cells isolated from the third trimester human placenta). Possibly due to the difference in the expression levels of the pluripotency‐related genes, induction of genes specific to the ectodermal tissues were more prominent in fPMSC than tPMSC after induced differentiation.
Role of HMGB1 in propofol protection of rat intestinal epithelial cells injured by heat shock
Authors: Tang Jing , Deng Peng , Jiang Yu , Tang Youqing , Chen Bin , Su Lei , Liu Zhifeng 
Published: August 2013

AbstractGut‐derived endotoxin and pathogenic bacteria may be important causative factors of morbidity and death during heat stroke. However, as the key component of intestinal mucosal barrier, the molecular mechanism of how intestinal epithelial cells are injured by heat shock is remains unclear. After rat intestinal epithelial cells (IEC‐6) had been exposed to heat shock, their viability was measured. Propofol, which plays an important role in anti‐inflammation and organ protection, was investigated to see how it affected viability under this stress. Changes of high mobility group box 1 (HMGB1) in IEC‐6 cells were measured with RT‐PCR and Western blot assay at transcription and translational levels, respectively. Ethyl pyruvate (EP), a specific inhibitor of HMGB1 that can inhibit the release of HMGB1 without affecting its intracellular synthesis, was also investigated. Heat shock significantly reduced the intracellular level of HMGB1, and propofol inhibit its reduction. Propofol protected the heat shock‐injured cells, at least partly through inhibiting the release of intracellular HMGB1 to reduce the direct or indirect cell damage caused by HMGB1. Pretreatment with high concentrations of EP also attenuated heat‐shock injury.
Polypeptide chain release factor eRF3 is a novel molecular partner of survivin
Authors: Xiao Ruilin , Gao Yan , Shen Quan , Li Cui , Chang Wenjuan , Chai Baofeng 
Published: August 2013

AbstractThe eukaryotic class II polypeptide chain release factor (eRF3) is an eRF1‐ and ribosome‐dependent GTPase involved in translation termination of protein biosynthesis. eRF3 is a multifunctional protein that is also involved in chromosomal segregation and cytokinesis during mitosis. Survivin is a member of the inhibitor of apoptosis protein (IAP) family that is involved in the organisation of spindle and cell apoptosis. Interaction between survivin and eRF3a‐F3 or eRF3b, encoded by the GSPT1 and GSPT2 genes, respectively, was confirmed using yeast two‐hybrid (Y2H) and pull‐down assays in vitro, and co‐immunoprecipitation in vivo. The domains involved in the formation of the survivin–eRF3s complex have been identified. The sites on survivin that interact with eRF3 are located in the baculovirus IAP repeat domain (residues 65–76), which forms a beta‐strand structure with an overall negative charge. The sites on eRF3 that interact with survivin were localised to the N‐terminal domain(NTD; residues 131–200). Cell localisation experiments indicate that both factors are in the nucleus, suggesting that they cooperatively function in nuclear processes.
Cytokine profiles, signalling pathways and effects of fluticasone propionate in respiratory syncytial virus‐infected human foetal lung fibroblasts
Authors: Seki Erina , Yoshizumi Masakazu , Tanaka Ryota , Ryo Akihide , Ishioka Taisei , Tsukagoshi Hiroyuki , Kozawa Kunihisa , Okayama Yoshimichi , Okabe‐Kado Junko , Goya Tomoyuki , Kimura Hirokazu 
Published: August 2013

AbstractTo examine cytokine production in response to RSV infection, we assessed the levels of 29 cytokines released from RSV‐infected human foetal lung fibroblasts. We also examined the relationships between the effects of fluticasone propionate and various signalling pathways in the cells. Twenty‐four hours after infection (1MOI), RSV‐infected cells released cytokines, for example proinflammatory cytokines (IL‐1β, IL‐6 and TNF‐α), anti‐inflammatory (IL‐1ra), Th1 (IFN‐γ, IFN‐λ1a, IL‐2 and IL‐12), Th2 (IL‐4, IL‐5, IL‐10 and IL‐13), granulopoiesis‐inducing (G‐CSF and GM‐CSF), eosinophil recruitment‐inducing (eotaxin and RANTES) and neutrophil recruitment‐inducing cytokines (IL‐8, IP‐10, MCP‐1 and MIP‐1α). Aberrant release of most was significantly suppressed by fluticasone propionate. Twelve hours after RSV infection, increased phosphorylation of Akt, p38 MAPK, ERK1/2 and IκB‐α was noted. Fluticasone propionate suppressed the phosphorylation of Akt, p38 MAPK, and ERK1/2, but not IκB‐α, in virus‐infected cells. TLR‐4 expression was unchanged in control and RSV‐infected cells, and TLR‐3 and RIG‐I expression was not detected. The results indicate that RSV infection induces aberrant production and release of certain cytokines through these signalling pathways in human lung fibroblasts. Overproduction and imbalance of these cytokines may be associated with the pathophysiology of RSV‐induced excessive and allergic inflammation.
Death inducing and cytoprotective autophagy in T‐47D cells by two common antibacterial drugs: sulphathiazole and sulphacetamide
Authors: Mohammadpour Raziye , Safarian Shahrokh , Sheibani Nader , Norouzi Saeed , Razazan Atefeh 
Published: August 2013

AbstractThe broad spectrum of the pharmacological effects of sulphonamide family of drugs motivated us to investigate the cellular mechanisms for anti‐cancer effects of sulphathiazole and sulphacetamide on T‐47D breast cancer cells. Fluorescent microscopy, flow cytometric analysis, caspase‐3 activity and DNA fragmentation assays were used to detect apoptosis. The distribution of the cells among different phases of the cell cycle was measured by flow cytometry. The expression of several genes with important roles in some critical cellular pathways including apoptosis, mTOR/AKT pathway and autophagy were determined by real‐time RT‐PCR analysis. Sulphathiazole and sulphacetamide induced anti‐proliferative effects on T‐47D cells were independent of apoptosis and cell cycle arrest. The overexpression of critical genes involved in autophagy including ATG5, p53 and DRAM indicated that the main effect of the drug‐induced anti‐proliferative effects was through induction of autophagy. This process was induced in two different forms, including death inducing and cytoprotective autophagy. Sulphathiazole treatment was followed by higher expression of p53/DRAM and downregulation of Akt/mTOR pathway resulting in death autophagy. In contrast, sulphacetamide treatment lowered expression of p53/DRAM pathway in parallel with upregulation of Akt/mTOR pathway promoting cytoprotective autophagy. The results indicated that autophagy is the main mechanism mediating the anti‐cancer effects of sulphathiazole and sulphacetamide on T‐47D cells. Alignment of the p53 and DRAM expression along with activation level of Akt survival pathway therefore determines the type of autophagy that occurs.
Intestinal stem cells – types and markers
Authors: Zhang Zhigang , Huang Jian 
Published: August 2013

AbstractIntestinal epithelium is a rapidly cycling tissue, always renewing every 4–5 days under normal conditions, which is maintained by intestinal stem cells (ISCs). Using the fluorescence labelling trace, ISCs can be divided into two different types: active intestinal stem cells (A‐ISCs) located in bottom of the intestinal crypt and the quiescent intestinal stem cells (Q‐ISCs) in the +4 position of the crypt. There is a complex signal regulation net between the ISCs and other intestinal cells, such as Wnt and Notch pathways. ISCs have an intimate relationship with the colorectal cancer (CRC). However, a deficiency of stem cells markers severely limits research on the biological characteristics of ISCs. We have reviewed several ISCs markers, including Lgr5, PHLDA1, Bmi1 and Lrig1. These markers have widely different biological functions, but also have a close relationship with cancers, especially CRC. Our hypothesis concerns the reasons for ISCs having two distinct types and why endless ISCs markers have emerged.
Insulin at a unicellular eukaryote level
Authors: Csaba György 
Published: August 2013

AbstractThe unicellular ciliate, Tetrahymena, has been the main model for studying the hormonal system of unicellular animals. Tetrahymena produce, store, secrete and take up insulin, the hormone being similar to that of mammals, both immunocytochemically and functionally. The plasma membrane and nuclear envelope of Tetrahymena have insulin receptors, which are structurally similar to the mammalian receptor, as it their binding capacity. The cell has also second messengers and signal pathways for insulin. Insulin influences the synthesis of other hormones. The first short encounter between the cell and insulin provokes the hormonal imprinting that alters the function of the cells and is transmitted to the progeny, and can persist for over 1,000 generations, in hormone binding, hormone content, phagocytosis, cell growth and movement. Insulin has a survival function in Tetrahymena and during stress insulin production and binding are elevated. Other protozoa also react to insulin, and the evolutionary aspects are discussed in this review since it is still not appreciated that the hormones are of great antiquity in the animal kingdom.
Higher propensity of Wharton's jelly derived mesenchymal stromal cells towards neuronal lineage in comparison to those derived from adipose and bone marrow
Authors: Balasubramanian Sudha , Thej Charan , Venugopal Parvathy , Priya Nancy , Zakaria Zubaidah , SundarRaj Swathi , Majumdar Anish Sen 
Published: August 2013

AbstractMesenchymal stromal cells (MSCs) derived from different tissue sources are capable of differentiating into neural and glial cell types. However, the efficiency of differentiation varies between MSCs derived from different tissues. We compared the efficiency of neural progenitor population generation between adipose (AD), bone marrow (BM) and Wharton's jelly (WJ) derived MSCs. MSCs isolated from the three sources were induced to form primary neurospheres using epidermal growth factor (20 ng/mL) and bFGF (20 ng/mL). The self‐renewal potential of the primary neurospheres was assessed by secondary neurosphere assay. Primary neurospheres were differentiated to neuronal lineage on fibronectin‐coated dishes. The neurospheres and the resulting differentiated cells were characterized by immunocytochemistry and the RT‐PCR analyses. We have also investigated the secretome profile of neuronal‐related growth factors using Ray biotech cytokine array. The results show that MSCs from the three sources can be induced to generate neurospheres and they expressed neural progenitor markers nestin, Sox2 and Pax6 transcription factors. When differentiated on fibronectin coated dishes in mitogen free culture conditions, the primary spheres from all three sources were able to generate neuron/glial ‐ like cells which expressed Nfl, Map2 and GFAP with varied efficiency. Self‐renewal potential of these progenitors was determined by secondary sphere formation. WJ‐ and BM‐derived neurospheres were able to self‐renew, while AD derived progenitors failed to do so. Comparison of the secretome profile suggested that WJ derived MSCs secrete more neurotrophic factors. The data suggest that human WJ derived MSCs can be induced to make neural progenitors with higher efficiency compared to BM and AD derived MSCs.
Aurora B kinase is required for cytokinesis through effecting spindle structure
Authors: Wu Qiang , Zhang Wei , Mu Tianhao , Song Tianqiang , Li Dengwen 
Published: August 2013

AbstractThe spindle, composed of microtubules and associated proteins, serves as major machinery for cell division. Aurora B is a chromosome passenger protein that has important functions in mitosis. Its dynamic distribution at mitosis goes along with spindle structure and dynamics. We used the siRNA technique to knockdown protein expression and immunofluorescence technique to follow Aurora B during mitosis. Aurora B regulates microtubule plus ends as mitosis progresses, including both kinetochore and polar microtubules. Interactions between Aurora B and polar microtubule plus ends lead to failure in cytokinesis and abnormal midbody structure. We think Aurora B may not only play a role as kinase, but regulate microtubule plus ends.
Novel interactions of GRP78: UPR and estrogen responses in the brain
Authors: Avila Marco Fidel , Cabezas Ricardo , Torrente Daniel , Gonzalez Janneth , Morales Ludis , Alvarez Lisandro , Capani Francisco , Barreto George E. 
Published: August 2013

AbstractGlucose‐regulated protein 78 (GRP78; 78 kDa) belongs to a group of highly conserved heat shock proteins (Hsp) with important functions at the cellular level. The emerging interest for GRP78 relies on its different functions, both in normal and pathological circumstances. GRP78 regulates intracellular calcium, protein shaping, endoplasmic reticulum (ER) stress and cell survival by an immediate response to insults, and that its expression may also be regulated by estrogens. Although these roles are well explored, the mechanisms by which GRP78 induces these changes are not completely understood. In this review, we highlight various aspects related to the GRP78 functioning in cellular protection and repair in response to ER stress and unfolded protein response by the regulation of intracellular Ca2+ and other mechanisms. In this respect, the novel interactions between GRP78 and estrogens, such as estradiol and others, are analyzed in the context of the central nervous system (CNS). We also discuss the importance of GRP78 and estrogens in brain diseases including ischemia, Alzheimer's and Huntington's disorders. Finally, the main protective mechanisms of GRP78 and estrogens during ER dysfunction in the brain are described, and the prospective roles of GRP78 in therapeutic processes.
Coexpression of recombinant adenovirus carrying GDNF and EDNRB genes in neural stem cells in vitro
Authors: Sun Nian‐Feng , Zhong Wen‐Yu , Lu Sheng‐Ai , Tian Yu‐Ling , Chen Jing‐Bo , Hu San‐Yuan , Tian Ai‐Ling 
Published: August 2013

AbstractGene therapy and nerve stem cells isolated from the developing human enteric nervous system (ENS) are significant. They may open the route for the cell therapy of Hirschsprung's disease (HD). We have constructed the recombinant adenovirus‐carrying glial cell line‐derived neurotrophic factor (GDNF) and endothelin receptor B (EDNRB) gene, and investigated the exosomatic coexpression in neural stem cells. The recombinant adenovirus Ad‐GE coexpressing GDNF and EDNRB gene was constructed by the AdEasy system and confirmed by the reverse transcription polymerase chain reaction (RT‐PCR) method. Expression of exogenous genes in neural stem cells after transfection was confirmed by the flow cytometry and real‐time fluorescence quantitative PCR. Fragments of pAd Track‐CMV‐GE were consistent with GDNF and EDNRB. The green fluorescence of the positive cells was followed by fluorescence microscopy at 24 h after NSCs had been transfected, reaching a peak at 72 h after transfection. Flow cytometry showed that the efficiency of transfection was 15.0, 23.6, and 25.4% at 24, 48 and 72 h respectively. Real‐time fluorescence quantitative PCR showed the expression levels of mRNA of GDNF and EDNRB in 48 and 72 h groups were obviously higher than that in 24 and 96 h groups. Recombinant adenovirus carrying GDNF and EDNRB genes are coexpressed in neural stem cells, which may offer the possibility of a novel approach to local combination gene therapy for Hirschsprung's disease.
Vinculin, cell mechanics and tumour cell invasion
Authors: Goldmann Wolfgang H. , Auernheimer Vera , Thievessen Ingo , Fabry Ben 
Published: August 2013

AbstractThe focal adhesion protein, vinculin, is important for transmitting mechanical forces and orchestrating mechanical signalling events. Deregulation of vinculin results in altered cell adhesion, contractility, motility and growth, all of which are important processes in cancer metastasis. This review summarises recent reports on the role of vinculin in cellular force generation and signalling, and discusses implications for a role of vinculin in promoting cancer cell migration in 3D environments.