| 1. |
- 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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| 2. |
- Aad, G., et al.
(författare)
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- 2012
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swepub:Mat__t (refereegranskat)
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| 3. |
- Aad, G., et al.
(författare)
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- 2013
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swepub:Mat__t (refereegranskat)
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| 4. |
- Aad, G., et al.
(författare)
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- 2011
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swepub:Mat__t (refereegranskat)
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| 5. |
- Liu, Yu-peng, et al.
(författare)
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应用酶热生物分析仪快速测量血糖的研究分析
- 2013
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Ingår i: Journal of Clinical and Experimental Medicine. ; 12:8, s. 579-581
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Tidskriftsartikel (refereegranskat)abstract
- Objective To compare the enzyme thermistor bioanalysis equipment with blood glucose meter which is analyzed by automatic biochemical analyzer, validate the correct rate of this equipment. The long-term aim of the project was to develop a prototype for a bedside monitor system for semi-continuous monitoring of the blood glucose concentration. This was made possible by using the special advantage of the thermal sensor technique in combination with the adjustment of flow. Methods 150 patients were collected in our department. The glucose level was determined using a single channel enzyme thermistor device. Then the results were compared with lab biochemistry analysator. Results There was no significant difference between these two methods (P>0.05). The measuring method with single channel enzyme thermistor device is a reliable method in determining blood glucose level. Single channel enzyme thermistor keep a coincidence with automatic biochemical analyzer. The assay cycle time was 3 minutes. Comparative analysis between our device and the clinical system device showed an excellent correlation for 150 patient blood samples. The device was stabile for hundreds of injections over a period of 45 days. Conclusion Determined glucose in whole blood using a single channel Enzyme Thermistor device can be used to assess the severity of an illness and for evaluating therapeutic efficacy. The ability of the device to analyze whole blood without any pretreatment makes it possible to develop real-time analysis.
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| 6. |
- Xie, Yan-Hua, et al.
(författare)
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Nuclear Factor-kappaB-Mediated Endothelin Receptor Up-Regulation Increases Renal Artery Contractility in Rats
- 2013
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Ingår i: Basic & Clinical Pharmacology & Toxicology. - : Wiley. - 1742-7843 .- 1742-7835. ; 113:6, s. 401-410
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Tidskriftsartikel (refereegranskat)abstract
- Increased renal artery contractility leads to renal vasospasm and ischaemia as well as kidney damage. This study was designed to examine the hypothesis that organ culture of renal arteries induces transcriptional up-regulation of endothelin type A (ETA) and type B2 (ETB2) receptors in the smooth muscle cells via activation of nuclear factor-kappaB (NF-B) and subsequently increases renal artery contractility. Rat renal artery segments were organ-cultured for 6 or 24hr to increase endothelin receptor-mediated contraction. To dissect molecular mechanisms involved in this process, inhibitors for NF-B signalling pathway (MG-132 and BMS345541), transcription (actinomycin D) and translation (cycloheximide) were used during organ culture. Endothelin receptors were studied using a sensitive myograph (functional contractility), real-time PCR (mRNA analysis) and immunohistochemistry (protein localization). Compared with fresh segments, contractile responses to endothelin-1 (non-selective endothelin receptor agonist) and sarafotoxin 6c (selective ETB receptor agonist) were significantly increased in the segments after 24hr of organ culture; ETB2 receptor-mediated maximal contraction increased from 2.7 +/- 0.5 to 135.3 +/- 5.1 (p<0.001), and potency (pEC(50)) of ETA receptor agonist increased from 8.20 +/- 0.04 to 8.72 +/- 0.07 (p<0.001). This was in parallel with increased corresponding mRNA and protein expression for ETA and ETB2 receptors. BMS345541, MG-132, actinomycin D or cyclohexamide, respectively, suppressed the up-regulation of ETA and ETB2 receptors. Immunostaining performed with specific antibody showed that IB was phosphorylated during organ culture. In conclusion, activation of NF-B mediates up-regulation of ETA and ETB2 receptors and subsequently increases renal artery contractility, which may contribute to renal vasospasm and ischaemia as well as kidney damage.
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| 7. |
- Xie, Yan-hua, et al.
(författare)
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Up-Regulation of G-Protein-Coupled Receptors for Endothelin and Thromboxane by Lipid-Soluble Smoke Particles in Renal Artery of Rat
- 2010
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Ingår i: Basic & Clinical Pharmacology & Toxicology. - : Wiley. - 1742-7843 .- 1742-7835. ; 107:4, s. 803-812
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Tidskriftsartikel (refereegranskat)abstract
- Up-regulation of G-protein-coupled receptors (GPCR) plays key roles in renal hypertension and cardiovascular disease pathogenesis. The present study was designed to examine if lipid-soluble cigarette smoking particles (DSP), nicotine and endotoxin (LPS), induce GPCR up-regulation for thromboxane A(2) (TP), endothelin type A (ETA) and type B (ETB) receptors in renal artery, and if intracellular signal mechanisms are involved. Renal artery segments of rats were exposed to DSP, nicotine or LPS, in organ culture for up to 24 hr. The GPCR-mediated contractions were recorded by using a myograph system. Expression of the GPCR was examined by real-time PCR and immunohistochemistry at mRNA and protein levels. Sarafatoxin 6c (S6c, selective ETB receptor agonist), endothelin-1 (ET-1, non-selective ETA and ETB receptor agonist) and 9,11-Dideoxy-9a,11a-methanoepoxy prostaglandin F-2a (U46619, a TP receptor agonist) induced contractions were significantly increased after the arterial segments exposed to DSP in a concentration-dependent (0.1-0.4 mu l/ml) manner, and S6c also induced a time-dependent contraction, compared to control (dimethyl sulfoxide). This was in parallel with enhanced mRNA expression for ETB receptor but not ETA and TP receptors, while increased protein expression for ETA, ETB and TP receptors was seen. The specific nuclear factor-kappa B (NF-kappa B) signal pathway inhibitor BMS345541 was applied to link DSP effects to the GPCR up-regulation. It totally abolished ETB receptor up-regulation, but not ETA and TP receptor up-regulations. Our results suggest that DSP transcriptionally up-regulated ETB receptor expression in rat renal artery via NF-kappa B signal pathways, whereas up-regulation of ETA and TP receptor-mediated contraction may involve post-transcriptional mechanisms.
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