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- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy
- 2012
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Ingår i: Autophagy. - : Informa UK Limited. - 1554-8635 .- 1554-8627. ; 8:4, s. 445-544
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Forskningsöversikt (refereegranskat)abstract
- In 2008 we published the first set of guidelines for standardizing research in autophagy. Since then, research on this topic has continued to accelerate, and many new scientists have entered the field. Our knowledge base and relevant new technologies have also been expanding. Accordingly, it is important to update these guidelines for monitoring autophagy in different organisms. Various reviews have described the range of assays that have been used for this purpose. Nevertheless, there continues to be confusion regarding acceptable methods to measure autophagy, especially in multicellular eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers or volume of autophagic elements (e.g., autophagosomes or autolysosomes) at any stage of the autophagic process vs. those that measure flux through the autophagy pathway (i.e., the complete process); thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from stimuli that result in increased autophagic activity, defined as increased autophagy induction coupled with increased delivery to, and degradation within, lysosomes (in most higher eukaryotes and some protists such as Dictyostelium) or the vacuole (in plants and fungi). In other words, it is especially important that investigators new to the field understand that the appearance of more autophagosomes does not necessarily equate with more autophagy. In fact, in many cases, autophagosomes accumulate because of a block in trafficking to lysosomes without a concomitant change in autophagosome biogenesis, whereas an increase in autolysosomes may reflect a reduction in degradative activity. Here, we present a set of guidelines for the selection and interpretation of methods for use by investigators who aim to examine macroautophagy and related processes, as well as for reviewers who need to provide realistic and reasonable critiques of papers that are focused on these processes. These guidelines are not meant to be a formulaic set of rules, because the appropriate assays depend in part on the question being asked and the system being used. In addition, we emphasize that no individual assay is guaranteed to be the most appropriate one in every situation, and we strongly recommend the use of multiple assays to monitor autophagy. In these guidelines, we consider these various methods of assessing autophagy and what information can, or cannot, be obtained from them. Finally, by discussing the merits and limits of particular autophagy assays, we hope to encourage technical innovation in the field.
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| 3. |
- Fransson, Moa, et al.
(författare)
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CAR/FoxP3-engineered T regulatory cells target the CNS and suppress EAE upon intranasal delivery
- 2012
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Ingår i: Journal of Neuroinflammation. - : Springer Science and Business Media LLC. - 1742-2094. ; 9, s. 112-
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND:Multiple sclerosis (MS) is an autoimmune disease of the central nervous system (CNS). In the murine experimental autoimmune encephalomyelitis (EAE) model of MS, T regulatory (Treg) cell therapy has proved to be beneficial, but generation of stable CNS-targeting Tregs needs further development. Here, we propose gene engineering to achieve CNS-targeting Tregs from naive CD4 cells and demonstrate their efficacy in the EAE model.METHODSCD4+T cells were modified utilizing a lentiviral vector system to express a chimeric antigen receptor (CAR) targeting myelin oligodendrocyte glycoprotein (MOG) in trans with the murine FoxP3 gene that drives Treg differentiation. The cells were evaluated in vitro for suppressive capacity and in C57BL/6 mice to treat EAE. Cells were administered by intranasal (i.n.) cell delivery.RESULTSThe engineered Tregs demonstrated suppressive capacity in vitro and could efficiently access various regions in the brain via i.n cell delivery. Clinical score 3 EAE mice were treated and the engineered Tregs suppressed ongoing encephalomyelitis as demonstrated by reduced disease symptoms as well as decreased IL-12 and IFNgamma mRNAs in brain tissue. Immunohistochemical markers for myelination (MBP) and reactive astrogliosis (GFAP) confirmed recovery in mice treated with engineered Tregs compared to controls. Symptomfree mice were echallenged with a second EAE-inducing inoculum but remained healthy, demonstrating the sustained effect of engineered Tregs.CONCLUSIONCNS-targeting Tregs delivered i.n. localized to the CNS and efficiently suppressed ongoing inflammation leading to diminished disease symptoms.
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| 5. |
- Lai, H., et al.
(författare)
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A novel isotropic conductive adhesive with Ag flakes, BN and SiC nanoparticles
- 2010
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Ingår i: 2010 International Symposium on Advanced Packaging Materials: Microtech, APM '10. - 9781424467563 ; , s. 49-53
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Konferensbidrag (övrigt vetenskapligt/konstnärligt)abstract
- Isotropic conductive adhesives (ICAs) with lower bonding temperature, higher resolution and environmental friendly have been used extensively in packaging process. In order to improve the electrical and thermal conductive properties of ICAs, two kinds of bimodal high temperature stable ICAs with matrix SHT6 and fillers with composition of macro silver flakes and boron nitride nanoparticles or macro silver flakes and silicon carbide nanoparticles were studied. In these two kinds of adhesives, the silver flakes were 75wt%, and the contents of nanoparticles were Owt%, 0.5wt%, 1.5wt%, 2.5wt%, 3wt%, 5wt% in weight. All the samples were cured at 150°C for 1 hour. SEM images and EDS results show the nanoparticles disperse randomly in the ICA. The electrical resistivity of these ICAs depends on the contents of silver flakes and is hardly affected by BN nanoparticles and SiC nanoparticles. The thermal conductivity of these ICAs increases firstly with the weight increase of the BN nanoparticles and SiC nanoparticles. And then it decreases when the content of the nanoparticles beyond a certain point. ©2010 IEEE.
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| 6. |
- Lai, Huaxiang, et al.
(författare)
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Effects of BN and SiC nanoparticles on properties of conductive adhesive
- 2010
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Ingår i: Proceedings - 2010 11th International Conference on Electronic Packaging Technology and High Density Packaging, ICEPT-HDP 2010; Xi'an; 16 August 2010 through 19 August 2010. - 9781424481422 ; :Article number 5582434, s. 235-239
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Konferensbidrag (refereegranskat)abstract
- Isotropic conductive adhesives (ICAs) are a type of interconnect material used more and more widely in computer, robot, mobile phone, LED and so on. Compared with traditional solders, isotropic conductive adhesives have better working plasticity, creep resistance and heat resistance. In particular, isotropic conductive adhesives are more responsive in high density packaging than solder, which determines the dominance of ICAs in high density packaging in the future. In order to improve the thermal conductivity of ICA with acceptable electrical conductivity, Ag flakers, BN and SiC nanoparticles were added into the matrix. The content of silver flakes was 75wt%, and the content of nanoparticles (BN or SiC) in the isotropic conductive adhesives were 0wt%, 0.5wt%, 1.5wt%, 2.5wt%, 3wt%, 5wt% in weight. The conductive adhesives were coated on the PCBs with stencil printing and fifty SR1206 chip components were mounted on a PCB using conductive adhesive. All samples were cured at 150°for 1h. Further research into the reliability of the above isotropic conductive adhesives after temperature & humidity and thermal-cycling was carried out to analyze the effects of BN and SiC nanoparticals on the properties of ICA. The condition of the temperature & humidity test was 85 ° /85%RH, 500h. The thermal-cycling test was -40 °∼125°, 500 cycles and the soaking time and ramping rate were 19min and ±15°/min. Changes to electrical resistance were used to estimate the reliability of the isotropic conductive adhesives in this study. The microstructure of the failure samples was observed using a Scanning Electron Microscope (SEM). The water absorption of all ICAs is the same after 94h temperature and humidity aging and the rate of water absorption is also the same during the aging. The ICA with 3% boron nitride nanoparticles and 75% micron silver flakes shows the best temperature and humidity reliability, with the fewest cracks on the interface between ICA and component. After 500h thermal cycling aging, the resistance reduces in the first 100h and maintains in a certain value after 332h cycled thermal. The samples have no wide cracks but a few small ones on the interface. © 2010 IEEE.
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| 8. |
- Rui, Manman, et al.
(författare)
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TiO2 Nanoparticles Functionalized Sn/3.0Ag/0.5Cu Lead-free Solder
- 2012
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Ingår i: Proceedings of IEEE CPMT 2012 International Conference on Electronic Packaging Technology & High Density Packaging. - 9781467316804 ; , s. 203-207
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Konferensbidrag (refereegranskat)abstract
- As the development of micro-systems moves towards higher speed and miniaturization, the requirement for interconnection density increases significantly. However, the use of conventional solders will be limited as the increasing I/O density lowers the pitches to very small size. Recently, there have been great developments in nano-composite solders. This paper investigated the influence of nano-titanium dioxide (TiO 2 ) of 99.9% purity on the wettability, micro-structural, melting and mechanical properties of Sn/3.0Ag/0.5Cu. The composite solder was prepared by mechanically mixing solder paste with TiO 2 nanoparticles for 30 minutes. The TiO 2 nanoparticles, with average diameter of 10 nm, were manufactured by precipitation. The solder paste was SAC305 Type 4. After reflow soldering the wetting angle was measured. The microstructure of the composite solders and TiO 2 was studied by means of scanning electron microscope (SEM). An optical microscope (OM) was employed to observe the fracture mode after shear test. A pull test was performed to investigate the shear strength of all samples with composite solders between two PCBs with Sn coating, both before and after thermal cycling (TC) with range between -40°C and 85°C. Differential scanning caborimetry (DSC) was adopted to analyze the melting point of composite solders. The results indicate that when the TiO 2 content increased from 0.5% to 2%, the wettability of the solder improved, which resulted in higher shear strength and better mechanical behavior. © 2012 IEEE.
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