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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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| 2. |
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| 3. |
- Hua, Kuo-Tai, et al.
(författare)
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N-α-acetyltransferase 10 protein suppresses cancer cell metastasis by binding PIX proteins and inhibiting Cdc42/Rac1 activity
- 2011
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Ingår i: Cancer Cell. - : Cell Press. - 1535-6108 .- 1878-3686. ; 19:2, s. 218-231
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Tidskriftsartikel (refereegranskat)abstract
- N-α-acetyltransferase 10 protein, Naa10p, is an N-acetyltransferase known to be involved in cell cycle control. We found that Naa10p was expressed lower in varieties of malignancies with lymph node metastasis compared with non-lymph node metastasis. Higher Naa10p expression correlates the survival of lung cancer patients. Naa10p significantly suppressed migration, tumor growth, and metastasis independent of its enzymatic activity. Instead, Naa10p binds to the GIT-binding domain of PIX, thereby preventing the formation of the GIT-PIX-Paxillin complex, resulting in reduced intrinsic Cdc42/Rac1 activity and decreased cell migration. Forced expression of PIX in Naa10-transfected tumor cells restored the migration and metastasis ability. We suggest that Naa10p functions as a tumor metastasis suppressor by disrupting the migratory complex, PIX-GIT- Paxillin, in cancer cells.
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| 4. |
- Kar, Sandeep, et al.
(författare)
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Health risks for human intake of aquacultural fish : Arsenic bioaccumulation and contamination
- 2011
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Ingår i: Journal of Environmental Science and Health. Part A. - : Informa UK Limited. - 1093-4529 .- 1532-4117. ; 46:11, s. 1266-1273
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Tidskriftsartikel (refereegranskat)abstract
- Aquacultural tilapia (Oreochromis mossambicus L.) and shrimp (Penaeus monodon L.) from groundwater-cultured ponds in southwestern Taiwan were analyzed to estimate arsenic (As) bioaccumulation and the potential health risk to human intake. Most of aquacultural ponds exhibited higher arsenic than maximum allowed concentrations (50 mu g L(-1)) in pond water of Taiwan. Arsenic levels in tilapia in Budai, Yichu and Beimen were 0.92 +/- 0.52 mu g g(-1), 0.93 +/- 0.19 mu g g(-1) and 0.76 +/- 0.03 mu g g(-1), respectively and in shrimp was 0.36 +/- 0.01 mu g g(-1) in Beimen. Total arsenic in tilapia is highly correlated (R(2) = 0.80) with total arsenic concentration of pond water. Total arsenic in fish showed high correlation with that in bone (R(2) = 0.98), head (R(2) = 0.97) and tissue (R(2) = 0.96). Organic arsenic species (DMA) was found higher relative to inorganic species of As(III) and As(V). The average percent contribution of inorganic arsenic to total arsenic in fish samples was 12.5% and ranged between 11.7 to 14.2%. Bioaccumulation factors (BAFs) for total arsenic in fish ranged from 10.3 to 22.1, whereas BAF for inorganic arsenic ranged from 1.33 to 2.82. The mean human health cancer risk associated with the ingestion of inorganic arsenic in the fish was estimated at 2.36 x 10(-4) +/- 0.99 x 10(-4), which is over 200 times greater than a de Minimus cancer risk of 1 x 10(-6). The mean human health hazard quotient associated with ingesting inorganic arsenic in the fish was 1.22 +/- 0.52, indicating that expected human exposure exceeds the reference dose for non-cancer health effects by 22%. These results suggest that the inhabitants in this region are being subjected to moderately elevated arsenic exposure through the consumption of tilapia and shrimp raised in aquaculture ponds.
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| 5. |
- Liu, Chia-Chuan, et al.
(författare)
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Biogeochemical interactions among the arsenic, iron, humic substances, and microbes in mud volcanoes in southern Taiwan
- 2011
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Ingår i: Journal of Environmental Science and Health. Part A. - : Informa UK Limited. - 1093-4529 .- 1532-4117. ; 46:11, s. 1218-1230
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Tidskriftsartikel (refereegranskat)abstract
- Fluid and mud samples collected from Hsiaokunshui (HKS), Wushanting (WST), Yenshuikeng (YSK), Kunshuiping (KSP), Liyushan (LYS), and Sinyangnyuhu (SYNH) mud volcanoes of southwestern Taiwan were characterized for major ions, humic substances (HS) and trace elements concentrations. The relationship between the release of arsenic (As) and activities of sulfate-reducing bacteria has been assessed to understand relevant geochemical processes in the mud volcanoes. Arsenic (0.02-0.06 mg/L) and humic substances (4.13 x 10(-4) to 1.64 x 10(-3) mM) in the fluids of mud volcanoes showed a positive correlation (r = 0.99, p < 0.05) except in Liyushan mud volcano. Arsenic and iron in mud sediments formed two separate groups i) high As, but low Fe in HKS, WST, and SYNH; and ii) low As, but high Fe in the YSK, KSP, and LYS mud volcanoes. The Eh(S.H.E). values of the mud volcano liquids were characterized by mild to strongly reducing conditions. The HKS, SYNH, and WST mud volcanoes (near the Chishan Fault) belongs to strong reducing environment (-33 to -116 mV), whereas the LYS, YSK, and KSP mud volcanoes located near the coastal plain are under mild reducing environment (-11 to 172 mV). At low Eh values mud volcanoes, saturation index (SI) values of poorly crystalline phases such as amorphous ferric hydroxide indicate understaturation, whereas saturation is reached in relatively high Eh(S.H.E.) values mud volcanoes. Arsenic contents in sediments are low, presumably due to its release to fluids (As/Fe ratio in YSK, KSP, and LYS sediment: 4.86 x 10(-4)-6.20 x 10(-4)). At low Eh(S.H.E.) values (mild to strong reducing environment), arsenic may co-precipitate with sulfides as a consequence of sulfate reduction (As/Fe ratios in WST, HKS, and SYNH sediments: 0.42-0.69).
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