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| 2. |
- 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. |
- Chen, Y. B., et al.
(författare)
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Inhibitory effects of endomorphin-2 on excitatory synaptic transmission and the neuronal excitability of sacral parasympathetic preganglionic neurons in young rats
- 2015
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Ingår i: Frontiers in Cellular Neuroscience. - : Frontiers Media SA. - 1662-5102. ; 9
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Tidskriftsartikel (refereegranskat)abstract
- The function of the urinary bladder is partly controlled by parasympathetic preganglionic neurons (PPNs) of the sacral parasympathetic nucleus (SPN). Our recent work demonstrated that endomorphin-2 (EM-2)-immunoreactive (IR) terminals form synapses with mu-opioid receptor (MOR)-expressing PPNs in the rat SPN. Here, we examined the effects of EM-2 on excitatory synaptic transmission and the neuronal excitability of the PPNs in young rats (24-30 days old) using a whole-cell patch-clamp approach. PPNs were identified by retrograde labeling with the fluorescent tracer tetramethylrhodamine-dextran (TMR). EM-2 (3 mu M) markedly decreased both the amplitude and the frequency of the spontaneous and miniature excitatory postsynaptic currents (sEPSCs and mEPSCs) of PPNs. EM-2 not only decreased the resting membrane potentials (RMPs) in 61.1% of the examined PPNs with half maximal response at the concentration of 0.282 mu M, but also increased the rheobase current and reduced the repetitive action potential firing of PPNs. Analysis of the current voltage relationship revealed that the EM-2-induced current was reversed at -95 +/- 2.5 mV and was suppressed by perfusion of the potassium channel blockers 4-aminopyridine (4 AP) or BaCl2 or by the addition of guanosine 5'-[beta-thioldiphosphate trilithium salt (GDP-beta-S) to the pipette solution, suggesting the involvement of the G-protein-coupled inwardly rectifying potassium (GIRK) channel. The above EM-2-invoked inhibitory effects were abolished by the MOR selective antagonist D-Phe-Cys-Tyr-D-Trp-Orn-Thr-Pen-Thr-NH2 (CTOP), indicating that the effects of EM-2 on PPNs were mediated by MOR via pre- and/or post-synaptic mechanisms. EM-2 activated pre- and post-synaptic MORs, inhibiting excitatory neurotransmitter release from the presynaptic terminals and decreasing the excitability of PPNs due to hyperpolarization of their membrane potentials, respectively. These inhibitory effects of EM-2 on PPNs at the spinal cord level may explain the mechanism of action of morphine treatment and morphine-induced bladder dysfunction in the clinic.
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| 4. |
- Ma, Jiang, et al.
(författare)
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Investigating hollandite-perovskite composite ceramics as a potential waste form for immobilization of radioactive cesium and strontium
- 2021
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Ingår i: Journal of Materials Science. - : Springer Nature. - 0022-2461 .- 1573-4803. ; 56:16, s. 9644-9654
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Tidskriftsartikel (refereegranskat)abstract
- Ceramic matrix containing zirconolite, hollandite, and perovskite phases is proposed as a potential host for HLW immobilization. Hollandite phase principally immobilizes Cs, while perovskite phase mainly immobilizes Sr. In this study, hollandite–perovskite composite ceramics are considered as a specialized waste form for immobilizing the separated Cs and Sr from HLW streams and synthesized by a solid-state reaction method at 1300 °C for 5 h. The phase compositions of the synthesized composites were characterized by XRD and BSE. The XRD results indicated that the as-prepared ceramics are composed of tetragonal hollandite Ba0.8Cs0.4Al2Ti6O16, cubic perovskite SrTiO3, alongside a lesser amount of TiO2. The BSE—EDX results confirm that Cs partitions into the hollandite matrix, while Sr incorporates into perovskite host with homogenous distribution. In addition, aqueous durability testing was carried out using the MCC-1 static leach test method. The normalized release rates of Cs and Sr in HP-3 sample (i.e., 75 wt% Ba0.8Cs0.4Al2Ti6O16 + 25 wt% SrTiO3) were < 10−2 g·m−2·d−1 after 42 days, exhibiting excellent chemical durability. These results indicate that the hollandite–perovskite ceramic matrix could be considered as a customized host matrix for immobilization of the separated Cs and Sr from HLW streams.
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| 5. |
- Wang, Lijing, et al.
(författare)
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Antioxidant High-Conductivity Copper Pastes Based on Core-Shell Copper Nanoparticles for Flexible Printed Electronics
- 2023
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Ingår i: Advanced Functional Materials. - : John Wiley & Sons. - 1616-301X .- 1616-3028. ; 33:26
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Tidskriftsartikel (refereegranskat)abstract
- As a nontoxic and cost-effective material, copper pastes have attracted great attention in both academia and industry. However, achieving the long-term stability of copper pastes remains challenging due to their susceptibility to oxidation. Therefore, stable copper nanoparticles with a Cu(0)-Cu(I) core-shell structure containing a surface passivation layer of formate ions-involved Cu(I) coordination polymers are developed. Based on the self-reducing nature of the passivation layer, the nanoparticle-based copper pastes can be sintered in <1 min, showing high electrical conductivity (220 000 S cm(-1)), mechanical flexibility, and long-term stability after sintering. The excellent properties of the developed copper pastes are even comparable with the ones of silver pastes. These stable copper pastes have broad applications in printed electronics (e.g., glucose sensors, RFID tags, and electromagnetic shielding films), showing great potential in the fabrication of flexible printed electronics.
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| 6. |
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| 7. |
- Erdem, Cemal, et al.
(författare)
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Proteomic screening and lasso regression reveal differential signaling in insulin and insulin-like growth factor I (IGF1) pathways
- 2016
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Ingår i: Molecular & Cellular Proteomics. - : Elsevier. - 1535-9476 .- 1535-9484. ; 15:9, s. 3045-3057
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Tidskriftsartikel (refereegranskat)abstract
- Insulin and insulin-like growth factor I (IGF1) influence cancer risk and progression through poorly understood mechanisms. To better understand the roles of insulin and IGF1 signaling in breast cancer, we combined proteomic screening with computational network inference to uncover differences in IGF1 and insulin induced signaling. Using reverse phase protein array, we measured the levels of 134 proteins in 21 breast cancer cell lines stimulated with IGF1 or insulin for up to 48 h. We then constructed directed protein expression networks using three separate methods: (i) lasso regression, (ii) conventional matrix inversion, and (iii) entropy maximization. These networks, named here as the time translation models, were analyzed and the inferred interactions were ranked by differential magnitude to identify pathway differences. The two top candidates, chosen for experimental validation, were shown to regulate IGF1/insulin induced phosphorylation events. First, acetyl-CoA carboxylase (ACC) knock-down was shown to increase the level of mitogen-activated protein kinase (MAPK) phosphorylation. Second, stable knock-down of E-Cadherin increased the phospho-Akt protein levels. Both of the knock-down perturbations incurred phosphorylation responses stronger in IGF1 stimulated cells compared with insulin. Overall, the time-translation modeling coupled to wet-lab experiments has proven to be powerful in inferring differential interactions downstream of IGF1 and insulin signaling, in vitro.
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| 8. |
- Gao, Xin-Fen, et al.
(författare)
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Three new species of Corydalis (Fumariaceae) from northwestern Sichuan China
- 2008
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Ingår i: Novon. - : Missouri Botanical Garden Press. - 1055-3177 .- 1945-6174. ; 18:3, s. 330-335
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Tidskriftsartikel (refereegranskat)abstract
- The jiuding Ridge (Sichuan, China) has been explored during the past several years by a team from chengdu Institute of Biology. Chinese Academy of Sciences, and three undescribed species of Corydalis DC. (Fumariaceae) have been revealed and are apparently endemic to this mountain. The new species were observed in forest (C. capitata X.F. Cao, Liden & Y.W. Wang), and alpine rocky limestone areas (C. schistostigma X.F. Cao, Liden, Y.W. Wang & Y.L. Peng). The two first species have their closest relatives in the Wolong-Balangshan District in the Wolong-Balangshan District (Wenchuan. Sichuan). Corydalis aeaeae differs from C. panada Liden & Y.W. Wang in its small size, few-flowered racemes, and broadly obtuse outer petals with low, short erests. Corydalis capitata differs from the C. flexuosa Franchet complex in the capitate racemes, small flowers with deeply serrate petals, and the peculiar, thin, strictly ecect lateral branches. corydalis schistostinma is unique in the C. curviflora Maximowicz ex Hemsley group, in the deeply cleft stigma and very forward-projecting crests to the inner petals, and possibly has iits affinities more to the north and northwest in the Hengduan Mountains.
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| 9. |
- Klionsky, Daniel J., et al.
(författare)
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Guidelines for the use and interpretation of assays for monitoring autophagy in higher eukaryotes
- 2008
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Ingår i: Autophagy. - : Landes Bioscience. - 1554-8627 .- 1554-8635. ; 4:2, s. 151-175
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Forskningsöversikt (refereegranskat)abstract
- Research in autophagy continues to accelerate,1 and as a result many new scientists are entering the field. Accordingly, it is important to establish a standard set of criteria for monitoring macroautophagy in different organisms. Recent reviews have described the range of assays that have been used for this purpose.2,3 There are many useful and convenient methods that can be used to monitor macroautophagy in yeast, but relatively few in other model systems, and there is much confusion regarding acceptable methods to measure macroautophagy in higher eukaryotes. A key point that needs to be emphasized is that there is a difference between measurements that monitor the numbers of autophagosomes versus those that measure flux through the autophagy pathway; thus, a block in macroautophagy that results in autophagosome accumulation needs to be differentiated from fully functional autophagy that includes delivery to, and degradation within, lysosomes (in most higher eukaryotes) or the vacuole (in plants and fungi). Here, we present a set of guidelines for the selection and interpretation of the methods that can be used by investigators who are attempting to examine macroautophagy and related processes, as well as by reviewers who need to provide realistic and reasonable critiques of papers that investigate these processes. This set of guidelines is 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 verify an autophagic response.
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| 10. |
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