Luminescent quantum dots (QDs)--semiconductor nanocrystals--are a promising alternative to organic dyes for fluorescence-based applications. We have developed procedures for using QDs to label live cells and have demonstrated their use for long-term multicolor imaging of live cells. The two approaches presented are (i) endocytic uptake of QDs and (ii) selective labeling of cell surface proteins with QDs conjugated to antibodies. Live cells labeled using these approaches were used for long-term multicolor imaging. The cells remained stably labeled for over a week as they grew and developed. These approaches should permit the simultaneous study of multiple cells over long periods of time as they proceed through growth and development. 相似文献
As a novel discovered regulated cell death pattern, ferroptosis has been associated with the development of Parkinson’s disease (PD) and has attracted widespread attention. Nevertheless, the relationship between ferroptosis and PD pathogenesis is still unclear. This study aims to investigate the effect of iron overload on dopaminergic (DA) neurons and its correlation with ferroptosis. Here we use nerve growth factor (NGF) induced PC12 cells which are derived from pheochromocytoma of the rat adrenal to establish a classical PD in vitro model. We found significantly decreased cell viability in NGF-PC12 cell under ammonium ferric citrate (FAC) administration. Moreover, excessive intracellular iron ions induced the increase of (reactive oxygen species) ROS release as well as the decrease of mitochondrial membrane potential in PC12-NGF cells. In addition, we also found that overloaded iron can activate cell apoptosis and ferroptosis pathways, which led to cell death. Furthermore, MPP-induced PD cells were characterized by mitochondrial shrinkage, decreased expression of glutathione peroxidase 4 (Gpx4) and ferritin heavy chain (FTH1), and increased divalent metal transporter (DMT1) and transferrin receptor 1 (TfR1) expression level. In contrast, Lip-1 and DFO increased the expression level of GPX4 and FTH1 compared to MPP-induced PD cell. In conclusion, we indicated that overloaded intracellular iron contributes to neurons death via apoptosis and ferroptosis pathways, while DFO, an iron chelator, can inhibit ferroptosis in order to protect the neurons in vitro.
Given the high occurrence of prostate cancer worldwide and one of the major sources of the discovery of new lead molecules being medicinal plants, this research undertook to investigate the possible anti-cancer activity of two natural cycloartanes; cycloartane-3,24,25-diol (extracted in our lab from Tillandsia recurvata) and cycloartane-3,24,25-triol (purchased). The inhibition of MRCKα kinase has emerged as a potential solution to restoring the tight regulation of normal cellular growth, the loss of which leads to cancer cell formation.
Methods
Kinase inhibition was investigated using competition binding (to the ATP sites) assays which have been previously established and authenticated and cell proliferation was measured using the WST-1 assay.
Results
Cycloartane-3,24,25-triol demonstrated strong selectivity towards the MRCKα kinase with a Kd50 of 0.26 μM from a total of 451 kinases investigated. Cycloartane-3,24,25-triol reduced the viability of PC-3 and DU145 cell lines with IC50 values of 2.226?±?0.28 μM and 1.67?±?0.18 μM respectively.
Conclusions
These results will prove useful in drug discovery as Cycloartane-3,24,25-triol has shown potential for development as an anti-cancer agent against prostate cancer. 相似文献
The membrane-anchored proteins of enveloped viruses form labile spikes on the virion surface, primed to undergo large-scale conformational changes culminating in virus-cell membrane fusion and viral entry. The prefusion form of these envelope glycoproteins thus represents an important molecular target for antiviral intervention. A critical roadblock to this endeavor has been our inability to produce the prefusion envelope glycoprotein trimer for biochemical and structural analysis. Through our studies of the GPC envelope glycoprotein of the hemorrhagic fever arenaviruses, we have shown that GPC is unique among class I viral fusion proteins in that the mature complex retains a stable signal peptide (SSP) in addition to the conventional receptor-binding and transmembrane fusion subunits. In this report we show that the recombinant GPC precursor can be produced as a discrete native-like trimer and that its proteolytic cleavage generates the mature glycoprotein. Proteoliposomes containing the cleaved GPC mediate pH-dependent membrane fusion, a characteristic feature of arenavirus entry. This reaction is inhibited by arenavirus-specific monoclonal antibodies and small-molecule fusion inhibitors. The in vitro reconstitution of GPC-mediated membrane-fusion activity offers unprecedented opportunities for biochemical and structural studies of arenavirus entry and its inhibition. To our knowledge, this report is the first to demonstrate functional reconstitution of membrane fusion by a viral envelope glycoprotein. 相似文献
Ohne ZusammenfassungVergl.Haorn (1946), sowieHaorn undNiggli (1946). Die anregung zu der vorliegenden Arbeit verdanke ich meinem verehrten Lehrer, Herrn Prof. Dr. E.Haorn, dem ich meinem herzlichen Dank aussprechen möchte für das Interesse, mit dem er sich meiner Arbeit widmete, für die zahlreichen Ratschläge und für die praktische Hilfe.Ebenso bin ich Herrn Prof.Hadorn für die Einführung in die Transplantationstechnik und für die Ueberlassang seiner Drosophilastämme grossen Dark schuldi.g 相似文献
A novel thermophilic anaerobic and microaerophilic bacterium (optimal growth in the presence of 5–10% O2), strain Nad S1T was isolated from the terrestrial hot spring of Hammam Sidi Jdidi, Nabeul, Tunisia. Cells were motile rods having a Gram-positive cell wall structure. Strain Nad S1T grew optimally at 55°C (range 37–70°C). Optimum pH for growth was 6.5–7.0. It was halotolerant growing with NaCl up to 7% (optimum concentration 1.5–3.0%). It grew chemoorganotrophically on various carbohydrates, organic-acids and amino-acids as energy sources, or chemolithotrophically on H2 using nitrate, as terminal electron acceptor. Beside oxygen (under microaerobic conditions) and nitrate, nitrite was also used. Nitrate was completely reduced to N2. No fermentation occurred. The genomic DNA G + C content was 41.8 mol%. Based on 16S rRNA gene sequence analysis, strain Nad S1T belongs to the Bacillaceae family within the class ‘Bacilli’. Because of its phylogenetic and phenotypic characteristics, we propose this isolate to be assigned as a novel genus and a novel species within the domain Bacteria, Microaerobacter geothermalis gen. nov., sp. nov. The type strain is Nad S1T (=DSM 22679T =JCM 16213T). 相似文献