l****j
发帖数: 70 | 1 看到一些文献说rapamycin能引起autophagy， 提高lysosome activity, 能促进
autophagosome fusion with lysosome,
另外又有文献说，rapamycin 只是部分抑制 mTORC1, 只能诱发autophagy, 对lysosome
不起作用，对autophagy的第二阶段也不起作用，
那么rapamycin的作用有没有统一性？还是不同细胞机制不一样？
请研究过rapamycin的朋友给点提示吧，谢谢！ |
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l****j
发帖数: 70 | 2 多谢回复 ！
那就是说rapamycin 还是多多少少会升高lysosome activity,
请问具体机制是什么？仅是通过抑制mTORC1就会升高lysosome activity吗？
有其他解释的途径吗？ |
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发帖数: 1 | 3 Lysosome stress反而会导致过度免疫激活。
: 氯喹好像就是抑制免疫反应再加上有一定抗病毒能力吧
|
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t******y
发帖数: 716 | 4 最近要做蛋白追踪，活细胞显影。但是没有亚细胞结构的荧光蛋白。如果哪位朋友有表
达mCherry，CFP等荧光蛋白偶联的ER，Lysosome或Golgi Marker的质粒，请和我联系。
不胜感激！ |
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m***6
发帖数: 424 | 5 Is there anyone who did lysosome staining?
I need good staining result. But with lysotracker could be done well.
Who could give some suggestions?
Thank you very much. |
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z*y
发帖数: 84 | 6 Have you tried lysotracker or not? You can try LAMP-1 or LAMP-2, both of
which are lysosome makers. |
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s******y
发帖数: 28562 | 7 endosome 和 phagosome主要是来源（膜上的蛋白）不同，导致后面的一
系列处理也不同
endosome 一般是由比较小的单分子结合在受体上引起的，一般需要
clathrin 来协助进入细胞
Phagosome是有比较大的复合体结合在细胞表面上引起的，不需要clathrin。
lysosome 是细胞内部包含着大量蛋白酶的泡泡，可以和其他泡泡融合之后来
降解里面的东西。 |
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y**u
发帖数: 7459 | 8 classic endocytosis, for cell surface receptors etc.:
clathrin-mediated endocytois --endosome (pH higher, sorting process here)--
lysosome(low pH, protein lysis)
phagocytosis, for large solid material, by forming larger PM ruffles ---
phagosome |
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r***e
发帖数: 2539 | 9 据我自己观察，加了rapamycin， lc3I/II两条带都变弱。
lysosome |
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T****u
发帖数: 424 | 10 rapamycin
Torin1
EBSS
HBSS
效果都不太一样。
drug浓度不一样，结果也不一样。
1. 用compound，一般compound都有各种off-target，纯在multiple effects
2. 简单地说autophagy有点笼统。
不同的marker表征的不一样，initionation or completion or others.
LC3-II先增加，但会recycle
这一点上不同drug，dosage不一样看到的结果也不一样。
还是看p62 or long-lived protein degradation
lysosome |
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w***r
发帖数: 709 | 11 扯淡一样的故事，关键是不是圈中人，我是捂裆派的。
Science. 2015 Jan 2;347(6217):83-6. doi: 10.1126/science.1258857.
Aging. Lysosomal signaling molecules regulate longevity in Caenorhabditis
elegans.
Folick A1, Oakley HD2, Yu Y2, Armstrong EH3, Kumari M4, Sanor L5, Moore DD6,
Ortlund EA3, Zechner R4, Wang MC7.
Author information
Abstract
Lysosomes are crucial cellular organelles for human health that function in
digestion and recycling of extracellular and intracellular macromolecules.
We describe a signaling role for ly... 阅读全帖 |
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q******g
发帖数: 3858 | 12 Chloroquine is a lysosomotropic agent that prevents endosomal acidification
[1]. It accumulates inside the acidic parts of the cell, including
endosomes and lysosomes.
This accumulation leads to inhibition of lysosomal enzymes that require an
acidic pH, and prevents fusion of endosomes and lysosomes.
Chloroquine is commonly used to study the role of endosomal acidification in
cellular processes [2, 3], such as the signaling of intracellular TLRs.
Moreover, Chloroquine inhibits autophagy as it ra... 阅读全帖 |
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j****g
发帖数: 406 | 13 谢谢老大的建议，下面是introduction的第一段。版面有限估计贴不了更多的了，如果
老大，或者是其他哪一位有兴趣的话我可以发到邮箱里。谢谢。
The balance between protein synthesis and degradation is important for
normal function and survival of cells. While the ubiquitin-proteasome
pathway is responsible for specific clearance of short-lived proteins, long-
lived proteins are delivered to lysosomes by autophagy, which is the major
inducible pathway for general turnover of bulk cytoplasmic components
including entire organelles such as mitochondria (Klionsky and ... 阅读全帖 |
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f******e
发帖数: 88 | 14 If the newborn screen (which includes the most common metabolic disease
alread) is negative. Please consider lysosomal storage disease.
Dried blood sample (same blood sample for newborn screen can be sent to
United Kindom for free test. You better ask the physcian give them a call
before you send the sample.
Please consider liver transplantation as doctor suggested.
Bless!
Biochemistry Department
Yorkhill Hospital
Yorkhill
Glasgow G3 8SJ
United Kingdom
Telephone: +44 (0) 141 ... 阅读全帖 |
|
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w*****t
发帖数: 179 | 16 您好，不知道我的背景能否审到这片文章。我曾经审过一些小杂志，希望能够对我争取
这次机会有所帮助。详细的CV和研究背景如下。非常感谢您的考虑。我的工作邮箱：
z****[email protected]
好像CV在这里粘贴很难看，不好意思。前一段时我的CV，后一段我的较详细的研究经历
。如有需要我更详细的背景情况，请跟我发信。
1
CURRICULUM VITAE
Full Name: Zhimin Wang
Address: Allegheny-Singer Research Institute, West Penn Allegheny, Health
System, South Tower, 320 East North Avenue, Pittsburgh, PA 15212-4772
Phone: 412-359-6153 (Office)
412-737-3777 (Cell)
E-mail: z****[email protected] w************[email protected]
EDUCATION
September, 2001- May, 2005 Ph.D., Molecular M... 阅读全帖 |
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w*****t
发帖数: 179 | 17 hello, thank you for providing the information. Please find my CV below and
my research experience. I have been revewing for over 10 journals for 14
times. My majour of PH.D is microbiology from Wuhan university. I have
published several papers concering virology and bacterial infection. So far
my work also foucse on salivary gland biofilm formation under LL37 and K4s4
treatment. If you need further information, please let me know. My email
address: z****[email protected]
Thank you for your time and c... 阅读全帖 |
|
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h*****4
发帖数: 36 | 19 师姐我来给你补充一下，
autophagy也是一个比较常见的降解通路，虽然最终经过lysosome降解蛋白质，但它不
等同于经典的lysosome降解(NH4Cl抑制的那种)。
常见的能block autophagy降解通路的药物有：3-MA, Bafilomycin A1 等等 还有能促
进的如Rapamycin等 可查阅相关文献。实际上autophagy最近这些年一直做得比较火热
。嘿嘿~
and |
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s*******n
发帖数: 37 | 20 Chloroqin increase PH of lysosome, and inhibit lysosome function, the last
step of autophagy. |
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k***r
发帖数: 100 | 21 I agree with Ankyrin, looks like endosomes and lysosomes (try EEA1 and some
lysozyme markers). Some times, truncated protein after translation it
misfolds and directly sent to lysosome for degradation. |
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m***g
发帖数: 197 | 22 怎么记得是CQ引起lysosome pH增加，而不能降解autolysosome，所以fre GFP积
累很多。
影响lyssome的pH同时，也block autophagosome和lysosome的fusion。
所以我觉得是两个方面都受阻。 |
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发帖数: 1 | 23 无意中看到一个老外写的，用了下面这篇文章里边介绍的使用荧光标签检控mitophagy。
Tools and techniques to measure mitophagy using fluorescence microscopy
DOI: 10.4161/auto.24001
FP-based biosensors for mitophagy
The field of FP engineering is accelerating with a plethora of new variants
described each year. These proteins show differen- tial properties, many of
which may provide useful tools to study mitophagy. For example, FP-based
biosensors such as a pH- sensitive mitochondrial-targeted FP chimera has
been recently developed to m... 阅读全帖 |
|
发帖数: 1 | 24 无意中看到一个老外写的，用了下面这篇文章里边介绍的使用荧光标签检控mitophagy。
Tools and techniques to measure mitophagy using fluorescence microscopy
DOI: 10.4161/auto.24001
FP-based biosensors for mitophagy
The field of FP engineering is accelerating with a plethora of new variants
described each year. These proteins show differen- tial properties, many of
which may provide useful tools to study mitophagy. For example, FP-based
biosensors such as a pH- sensitive mitochondrial-targeted FP chimera has
been recently developed to m... 阅读全帖 |
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g****t
发帖数: 12 | 25 备考过程中看了很多前辈的考经，医学职业版是一个好地方，为表示支持和感恩，自己
抽点时间简单分享一下我的备考过程：
复习资料：FA 2013 + Pathoma(online lecture and hardcopy of the notes) + UW
qbank + Kaplan qbank (只完成60-70%）+ Goljan audio ( Goljan high yield notes
只迅速过了1遍）
Step 1 2015年1月20日，2月11日收到成绩242。
Time line:
2013年底艰难的看完第1遍First aid, 同时完成ECFMG报名，平时主要忙于做实验，发
表文章和开会，有空的时候看了一点youtube里面能搜到的kaplan lecture, 主要是
biochemistry和pharmacology；这个过程主要是熟悉词汇，对FA有个整体印象，之后复
习就不用太频繁的查单词。
2014年初为毕业后申请OPT，3月底4月中旬看完第2遍First aid；过了一遍Goljan
pathology notes, 4月买了UW qbank， 4月到7月... 阅读全帖 |
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w********2
发帖数: 632 | 26 Publication Cover
Journal
The Journal of Slavic Military Studies
Volume 19, 2006 - Issue 2
Journal homepage
186
Views
0
CrossRef citations
3
Altmetric
Original Articles
British “Lend-Lease” Tanks and the Battle for Moscow, November–December
1941—A Research Note
Alexander Hill
Pages 289-294 | Published online: 21 Sep 2006
Download citation https://doi.org/10.1080/13518040600697811
Select Language▼
Translator disclaimer
Full Article Figures & data Citations Metrics Reprints & Permis... 阅读全帖 |
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s******n
发帖数: 518 | 27 既然你关心农药，那就推荐一个孟山都的roundup的危害的文章：
https://www.ecowatch.com/15-health-problems-linked-to-monsantos-roundup-
1882002128.html
Food
Guest Contributor
Jan. 23, 2015 11:44AM EST
15 Health Problems Linked to Monsanto's Roundup
Monsanto invented the herbicide glyphosate and brought it to market under
the trade name Roundup in 1974, after DDT was banned. But it wasn’t until
the late 1990s that the use of Roundup surged, thanks to Monsanto's
ingenious marketing strategy. The strategy? Genetically engineer ... 阅读全帖 |
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p****n
发帖数: 9263 | 28 another 女老印造假
RETRACTED: VMA21 Deficiency Causes an Autophagic Myopathy by Compromising V-
ATPase Activity and Lysosomal Acidification
Reason: Our paper reported the identification of mutations in the
gene VMA21 in patients with X-linked Myopathy with Excessive Autophagy (XMEA
) and characterized the molecular mechanisms underlying the disease
phenotype. Many of the figure panels in the paper summarize data from
multiple experiments. We have now detected a number of errors in these
panels |
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s********n
发帖数: 26222 | 29 方舟子在此文中说“虫子不吃的， 人能吃吗”是中国人的质疑， 而美国人是不会问这
样的问题的， 这显然是骗人的。见下文和其中引用的文献， 其中不少是美国人的科研
。
"Comments on the human health impact of Bacillus thuringiensis toxin gene
product in genetically modified crops"
EPA Review Docket Number OPP-00678B
Professor Joe Cummins
Professor Emeritus of Genetics
University of Western Ontario
I have studied the Environment Protection Agency (EPA) document "Bt Plant-
Pesticides Biopesticides Registration Action Document B. HUMAN HEALTH
ASSESSMENT" and comment in the following ... 阅读全帖 |
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m**c
发帖数: 7349 | 30 He began his prize-winning work in 1988, looking at autophagy in yeast. This
is a well-studied organism, often used by biologists to examine fundamental
cellular processes.
Unfortunately, its cells are small and to start with Dr Ohsumi could not
easily distinguish the autophagosomes within them under a microscope. He
dealt with this difficulty by disabling the manufacture of the lysosomic
digestive enzymes.
That meant autophagosomes were no longer destroyed, and accumulated to the
point where th... 阅读全帖 |
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发帖数: 1 | 31 不可能抗病毒。相反会降低macrophage, neutrophil的清除能力。 |
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w*******2
发帖数: 2199 | 32 操，你们千老就鸡巴会捅词1
氯喹已经在体外证实能抑制病毒了
你悲愤也没有用 |
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发帖数: 1 | 33 我表示高度怀疑。十个字
: 操，你们千老就鸡巴会捅词1
: 氯喹已经在体外证实能抑制病毒了
: 你悲愤也没有用
|
|
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发帖数: 1 | 35 氯喹好像就是抑制免疫反应再加上有一定抗病毒能力吧 |
|
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发帖数: 1 | 37 体外有个鸡巴用处。用食盐体外也能抑制病毒
盹盹盹 |
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a****c
发帖数: 339 | 38 用于绿卡申请。
关键字：cell biology/membrane trafficking/lysosomal function
曾经给3-4分的杂志独立审稿4次，帮老板审的10来次吧。
需要先看CV的请站内。先行谢过 |
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l*********6
发帖数: 58 | 39 谢谢美眉的信息！现在等待genetic的结果，确定分型，如果是I型治疗就有希望了，祈
祷祝福宝宝！
deficiency of beta glucocerebrosidase in lysosome and there are accumulated
glucocerebroside. There might be enzyme replacement therapy for this disease
. Please check the following link. |
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x***x
发帖数: 416 | 40 More common: 可能的常见的问题:
1. Glaucoma;
2. after central retinal vein or artery occlusion;
3. ischemic optic neuropathy;
4. chronic optic neuritis;
5. chronic papilledema;
6. compression of the optic nerve, chiasm, or tract by a tumor or aneurysm;
7. traumatic optic neuropathy
这些对应的诊断应该不难, 需要:VF, NMR, Dilated Fundus pictures, imaging, etc.
Less common:
8. Syphilis, 查CRP等梅毒指标,
9. retinal degeneration (e.g., retinitis pigmentosa), Dilated Fundus picture
,一看就知道了,
10. toxic/metabolic optic neuropathy, 有... 阅读全帖 |
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a****c
发帖数: 339 | 42 先行谢过。关键字：cell biology/membrane trafficking/lysosomal function |
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w*****t
发帖数: 179 | 43 Thank you for providing the review opportunity, I have been working in the
molecular biology for over 15 years and published some papers in journals
like Journal of Cell Biology, Journal of General Virology et al., . Also I
have reviewed over 20 manuscripts for 10 journals. Hopefully these
experiences will help me to catch this opportunity. If you need further
detail information regarding my qualification, please let me know (zwang1@
wpahs.org). Please find my CV below. Thanks a lot.
Zhimin Wang... 阅读全帖 |
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a***o
发帖数: 104 | 44 Keywords: lysosomal fusion; optogenetics; ion channel; calcium
请PM你的英文姓名，单位与邮箱以及发表过的相关文章, 我会向编辑推荐,不过不保证
成功率. |
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l******d
发帖数: 2186 | 45 应该是lysosome的自溶，给记者们转来转去，就变成这样了... |
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w***7
发帖数: 1637 | 46 bless
你朋友的小孩实在哪儿被确诊的？这个病非常稀有，国内居然有人知道，很奇怪。
这个病是由于lysosome 里面一种叫acid alpha-glucosidase 突变引起的。除了你所说
的ERT外，吃protein-enriched nutrition with supplementation of Ala and leu 也
会有治疗效果。 |
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d*****r
发帖数: 2583 | 47 ☆─────────────────────────────────────☆
nmda (SingingMouse) 于 (Mon Feb 2 16:26:06 2009) 提到:
表达一个和膜有关的GFP fusion protein然后看他的localization,同时用各种oganell
marker去染co-localization,看这个fusion protein定位或者聚集在哪个oganell,这
个方法和结果有多大的可信度? 有人说完全不可信,任选一个oganell marker,总能找到
一个细胞co-localized;有人说所有这个protein到过的地方都应该能找到co-
localization,所以ER,Golgi,endosome,lysosome都应该有co-localization. 如果这样
,那到底应该如何胞内定位这个protein?
☆─────────────────────────────────────☆
n158 (土人) 于 (Mon Feb 2 17:12:41 2009) 提到:
I think it i |
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s******y
发帖数: 28562 | 48 Promega, TnT T7 coupled transcription/translation kit
Depending on what degradation system was responsible. In vitro translation
system also contain ubiquitin and proteasome. However, it does not have
lysosome or golgi (which also contains protein degradation machinery).
You can add 10uM MG132 in the in vitro translation system (or your 293T cell
culture medium) to inhibit the proteasome.
的系统更容易降解我的蛋白？
Yes. If you want, you can try bacterial in vitro expression system. BUT, it
won't contain the |
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a******u
发帖数: 211 | 49 Sci Signal. 2009 May 19;2(71):ra23.
TRPM2 functions as a lysosomal Ca2+-release channel in beta cells.
Lange I, Yamamoto S, Partida-Sanchez S, Mori Y, Fleig A, Penner R.
Please send it to e*********[email protected]
Thanks so much |
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t*x
发帖数: 1065 | 50 呵呵，谢谢详细的回复！
那我就用RIPA buffer试一下
不过像你说的，如果这种现象正常的话，估计换了buffer也应该差不多的趋势。:)
刚开始的时候没有用sonication，只是0.5%triton X-100 30min 冰浴裂解
后来听别人建议，加了sonication,发现跑完westernblot，结果很一致，都是处理12小
时之后，在镜下观察发现vacuole都消失差不多了，才看到lc-3II的增加
呵呵，我以为是开始时lc3-II在vacuole膜上镶嵌，没有裂解下来，而时间长了
autophagosome跟lysosome结合吞噬，lc-3II被从膜上释放出来，成为cytosolic形式才
能被我检测到呢，所以一直在找比较强的裂解膜上蛋白的方法。
你这样说，我就比较放心了，恩，非常感谢！
western |
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