Idiotic Business Behavior by Large Banks, A Lesson to Be Learned

When I run my business we run into "situations" or more precisely, difficult customers, when you really want to rather not have that customer instead of doing all the extra work it requires to maintain that customer. But never in my clear mind did we imagine going through something that I just encountered at Bank of America, one of our banks for 20 years. It was a simple situation, they set us up for a type of checking account that somehow stipulated that if we did not maintain $10,000 balance we would be charged a $20 monthly maintenance fee. Since I'd never paid monthly fee for our personal or the business accounts of allelebiotech.com, I probably did not bother to ask why the change. The things got tough in late 2008 to early 2009, I had to scramble all personal funds to not lay off employees, who were going through their own personal difficulties especially the younger ones who were paid by hourly.

Now things are better, I get to pay myself a salary again and pay more attention to my own bank accounts, and one of the things I noticed was that I was being charged a $20 fee at some point during the last 12 months. I asked for a refund for a long term customer's sake, got rejected; then I asked to be removed of any such future fees because my business banks always offer unconditional free checking to all my employees, and got rejected again because "the fee was charged correctly". They did offer a solution, go online open a new checking account since all new accounts opened online will be free of fees.

I did open a new account online, and all its online banking features are exactly the same as the checking account I have been using for 20 years, and online for maybe 10 years. Now I have to redo all my automatic payer and payee accounts, direct deposit, transfer routes... to save $20 a month using exactly the same service. If BOA meant to get a protocol right to charge fees while being able to attract new customers through free checking and encouraging online users to save labor costs, they succeeded, I understand their unspoken business reasons. But it they meant to bother customers to a degree that they would rather leave, they have sure succeeded in a unique way, by which business sense no longer aligns with common sense. The inflexibility they demonstrated through their lower level representatives is quite impressive, I would not allow such handling in my own company. It's simple, the cost to attract and maintain a loyal customer like myself, for 20 years, would be so much higher than somehow partially refunding the $20 fees, or a simple management check to remove future fees without forcing me to open a new account.

I see this as a business real situation lesson, and I know I will be fine doing all my business at my business banks. I am just not quite sure if I should also get rid off all my BOA shares, that may seems like an emotional decisions rather then business though...Oh I also will write to my congressmen and my president not to support more bailout money, they encourage bad business attitude.

傅国涌：林昭正悄悄回到我们的生活中

Note from the blog admin: It is still this Blog's policy to keep repost, retweet to the minimum and maintain this site as a source of original contents. However, this article meets the requirement of being rare in content and unique in style. The topic also suites perfectly the date of today, Christmas Eve 2009.

在鲁迅的故乡绍兴，2009年的中考语文试卷中出现了“林昭”的名字，这是一道成语运用的选择题：

　　下面句子中，加点的成语运用不恰当的一项是( )

　　A.他心高气傲，总喜欢(妄自菲薄)他人，结果可想而知，没有人原意同他打交道，他成了大海里的一叶孤舟。

　　B.从太空回眸我们这颗生存的星球，在(变幻莫测)的白去的飘忽中，它总是散发着一种浅蓝色的绚丽，谁也说不清那究竟是大海还是气晕的颜色。

　　C.林昭曾说：“我们的血是再鲜艳不过，而且是再灿烂不过的墨水，人世间其它一切墨水在这样的墨水面前统统不免(黯然失色) 。”

　　D.凭着健壮的体魄，你可以支撑起一方蔚蓝的天空；凭着旺盛的精力，你可以开垦出一片神奇的土地；凭着巨大的潜力，你可以变得出类拔萃，令人(刮目相看)。

　　林昭，以这样的方式出现在中学生的眼中，也许还是第一次，尽管他们中绝大多数不知道林昭是谁，在匆忙的考试中也顾不上去追问，但，林昭这个名字与他们的生命就这样相遇了，而且是在一次重要的考试当中。我最初是从一个朋友的博客看到这个消息，然后顺藤摸瓜找到了出处，名叫《归忆江天发浩歌》的博客上6月 20日发表的博文《中考卷里的林昭》。

　　这篇博文在我们周围的朋友中引起了小小的惊动，因为林昭的缘故，大家都想认识这个博客的主人。确实，在被杀戮40年后，林昭正悄悄地回到我们的生活中。

　　记得去年冬天，几个朋友一同去乌镇游玩的路上，我偶然在一个朋友的车上听到播放的音乐竟是林昭的《普罗米修斯受难的一日》，这是云南一个摇滚乐队的作品，主唱者刘涛，他们的乐队叫做“腰乐队”，都很年轻，他们把林昭的诗谱曲、演唱，并制成了光盘，在网上流传，我们听了都很感动。通过唱片上留下的电子信箱，我与刘涛联系上了，才知道在西南边陲有这样一群怀抱理想主义情怀的民间音乐人。

　　也是去年，我去湖北一所中学给高一学生做一个《科学家的人文视野》讲座，讲座结束时，我接到许多学生的纸条，有好几个不约而同地问的是林昭、“红楼”的林姑娘，其中一个还将毛泽东和林昭放在一起问：您认为毛泽东是个什么人？对于林昭的人生您有何看法？他们希望多了解一点林昭的历史，这不免让我有些意外，这些90后的学生也开始在探寻真相了。在林昭身后 40年，她的英灵不仅没有消散，而且带着更大的不可战胜的力量回来了，回到我们的中间，与我们这块土地同在。

　　去年5月1日，《南方周末》以《读林昭十四万言书》为题发表我为纪念林昭被杀40周年而写的文章，我曾接到编辑转来一位叫方震的读者来信，他说，读此文“泪如泉涌，泣不成声”，并写下一诗《哀林昭》，请编辑转给我：檄文读罢泪沾襟，漆夜中华一点萤。天下男儿当有愧，林昭青史独留名。

　　林昭已成为民族记忆中一个不可替代的符号，每一个良知未泯的国人一旦了解她的生平、思想和殉难之惨烈，无不对她肃然起敬。80多岁的律师张思之先生将这自己在林昭墓前的留影视为珍贵，90多岁的学者刘绪贻先生惦念着林昭，更多的年轻人把林昭视为20世纪中国精神史上的偶像，“黑暗时代”的一道亮光，中国的基督徒以拥有林昭这样一位殉道的姊妹而深感欣慰。不仅年年的清明节和4月29日的林昭祭日，就是在平日，苏州城外灵岩山上前往扫墓的人也是络绎不绝，生财有道的当地村民甚至形成了为林昭墓带路的小小“产业”，向四面八方前来为林昭扫墓的人收取带路钱，这已经是灵岩山的一道有点煞风景的“风景”。即使去年40周年祭前夕，当局在林昭墓前的树上安装了意为恐吓的摄像头，也没有阻止人们走近林昭的脚步。尽管网上的“林昭小组”、“林昭群”都被解散了，关于林昭的文字也常常被屏蔽。但是，这一切都没有能挡住林昭回到我们生活当中——

　　在山东，一位年轻的女教师海虹，以林昭为原型，写了一部题为《中国红豆词》的长篇小说，2008年6月由中国国际文化出版社出版，本来小说的主角就叫林昭，只是出版时未能通过，无奈地改成了“林萍”，整个故事几乎没有虚构，就是以文学的形式重叙了林昭的生平。

　　在南京，一位年轻的女记者赵锐，为林昭的故事所感动，完成了一本《祭坛上的圣女——林昭传》，已在台北秀威信息公司出版，并于不久前凤凰新华集团在南京举办的“2009年中国南京秋季馆藏图书展销会”
公开陈列展销。

　　在北京，一位年轻的诗人朵朵，写了一出以林昭为主题的独幕剧。也是在北京，以“祭园守园人”自居的朱毅先生，有意倡议为林昭建一个铜像，最好当然是立在北大未名湖畔，退而求其次，则可以放在林昭墓前……

　　戴镣铐的泉，她将永葆青春和甘甜
　　戴镣铐的血，已经自由地流在她体外
　　……受难是她每日的生活，受难是她自愿的选择。
　　……
　　她用盛大的爱来勾兑盛大的苦难
　　在一切黑暗之上，“爱”站立起来
　　“爱”举起的火把永不熄灭
　　……
　　戴镣铐的泉，她将永葆青春和甘甜
　　传薪火的人，她已经来到我们身边

　　诗人三春晖在林昭祭日写下了这首《戴镣铐的泉》，镣铐锁不住泉的涌流，血的涌流，镣铐也锁不住爱的涌流。向死而生的林昭，留下的不是恨的种子，她撒布的是爱和宽恕，那正是一个古老民族所缺少的因子，那是从基督而来的，是通向一个新时代的基石。

　　谷雨过后　大地上的桃花一齐败了
　　落满我的掌心
　　我不是刽子手
　　但我却攥出了一手 淋漓的鲜血
　　……
　　谷雨过后 我从潮湿的江南起身
　　去寻找一支遗失的火炬
　　在石头中　我看见你的名字
　　泼洒着太阳的热情

　　这是诗人涂国文为纪念林昭殉难四十周年写的诗《谷雨过后》，2009年9月，浙江文艺出版社出版他的诗文集《苏小墓前人如织》就收入了此诗，林昭逃过了编辑的“火眼金睛”，这在向来以自律、不出“差错”为金科玉律的浙江出版界，真是一件不容易的事，也许编辑、审稿者压根没有听说过林昭这个名字，所以才侥幸漏网。但是，这样的漏网总是好事，毕竟可以使更多的人听说林昭的名字。

　　在遥远的星空，林昭，正默默地注视着她深深热爱的这片土地。如果能够，她当然不喜欢用血写诗，虽然“人世间其它一切墨水在这样的墨水面前统统不免黯然失色”。她是那么热爱生活，她根本不是政治中人，她不喜欢政治，厌恶那些讲权谋、讲成败的政治，她所争的是是非，是人格，是尊严，这一切在她看来，都是一个人生活下去不可缺少的因素，这一切一旦全部丧失了，那生活当中还剩下什么？她可以为此而死。林昭之死，不只是为她自己争人格，也是为全体国人争人格。今天，她之所以悄无声息地回到我们的生活中，是因为这个时代，我们仍然需要为自己争人格，林昭就是一个最好的本土榜样，她是中国的朋霍费尔，她走过的这条道路已经表明，她已成为古老民族争人格进程中最丰厚的精神资源，因为她提供的不仅有无比锐利的武器，而且有爱的滋润和宽恕敌人的胸怀。幽暗的星空，因着她那双闪烁不定的眼睛，而不再虚空，不再无助，不再绝望。

周末看关于贪官下马新闻随想

最近一年内第三个下马居贪省政协（！）主席的背后故事中出现吴姓高级官员, 感叹政协腐败亦能如此之余，忽然想起大陆出来的人中对于进入官场的热忱。可能一辈子没当过官，也不知当官为何味的普通学术人中亦行此风。认识一位，某次因为有可能有一机会被那位吴姓“有前程”官员接见而兴奋不已，愿千里飞回以“科技”为敲门砖自荐。 可惜该官未成第四代。另一位则以自家人中能有人以学术而进官场（有现成最大先例为榜样）为长远计划，听说某个干部子弟来附近学校上学，即放下长辈、精英架子，主动要为该公子提供关照，尽管黑官是我们作为青年学生时要用毕生理想和一腔鲜血反对的社会腐朽，尽管贪官公子多半儿是在消耗包括自己父母在内的百姓的民脂民膏。不知那个官员到底是谁但好象没有起到什么作用，也许是因为想提供关照者最后也没能关照上，也许该官员没作用力。想起这些并没有低视把当官作为理想追从的人的感觉，微笑之余，忽有一种对生活、思想自由的更多的珍惜，对为人而独立于天地之间无精神束缚的豪爽，和对我在人生选择中有极大启迪的父亲的怀念。

mTFP1 is an excellent FRET donor

Because of its excitation and emission wavelength, sharp excitation and emission peaks, high quantum yield, and exceptional photostability, mTFP1 has always been considered a very good Forster resonance energy transfer (FRET) donor (1). More recently, several groups have investigated the use of mTFP1 in various FRET experiments and imaging modalities and have shown that mTFP1 is indeed one of the best choices (2, 3, 4).

In one recent publication, Padilla-Parra et al (2) tested a number of different FRET couples to determine which was the best for fluorescence lifetime imaging (FLIM)-FRET experiments, and found that the mTFP1-EYFP pair was by far the best pair for FLIM-FRET. This group also confirmed that the fluorescence lifetime decay of mTFP1 fits well to a single exponential, and that the time constant for this decay is unaffected by photobleaching, making mTFP1 an excellent choice for any kind of fluorescence lifetime imaging applications, including FLIM-FRET. This group also notes that it is likely that the use of Venus or mCitrine variants in place of EYFP would improve the performance of this FRET pair even further.

In a mathematical analysis of the potential FRET efficiency of mTFP1 with Venus YFP, Day et al. (3) showed that compared with Cerulean (currently the brightest cyan Aequorea GFP variant), one can expect up to 17% better FRET efficiency using mTFP1. This group went on to characterize the mTFP1-Venus pair in live-cell FRET and FLIM-FRET experiments and showed that it worked as predicted in both cases. They also note that mTFP1 has superior brightness and photostability when compared to Cerulean in live cells, which is consistent with all in vitro data reported previously (1). In a related paper, Sun et al. (4) demonstrated that mTFP1 is also an excellent FRET donor for the orange fluorescent protein mKO2.

Together, these recent independent studies confirm that mTFP1 among the best options when choosing a fluorescent protein as a FRET donor. With its proven track record of successful fusions, mTFP1 is also an excellent all-around performer that will enhance almost any live-cell imaging experiment.

(1) Ai et al., (2006) Biochem. J. 400:531-540.
(2) Padilla-Parra et al., (2009) Biophys J. 97(8):2368-76.
(3) Day et al., (2008) J Biomed Opt. 13(3):031203.
(4) Sun et al., (2009) J Biomed Opt. 14(5):054009.

AlleleBlog Admin, by Nathan Shaner

Video of the month (NEW!): Protein Expression Systems on youtube (http://www.youtube.com/watch?v=n81orbUebsQ) and at our protein expression page.

Discount of the week (Dec 14-20): 15% off Phoenix Retrovirus Expression System 2.0 (with selection medium provided)

New product(s) of the week: 48 fluorescent protein fusions on ready-to-infect virus that get into primary mammalian cells as subcellular markers (http://www.allelebiotech.com/shopcart/index.php?c=197&sc=34), 20 infections, only $249 for a limited introduction time.

ASCB Abstract: Increased rate of reprogramming of induced pluripotent stem cells using high-titer lentiviral vectors encoding multiple cell growth and survival regulatory genes

Objective: Differentiated cells can be reprogrammed into induced pluripotent stem cells (iPSC) with enforced expression of multiple transcription factors. We aim to improve the reprogramming efficiency using high titer lentiviral vectors encoding additional cell growth and survival regulatory genes.

Methods: Lentiviral vectors encoding multiple cell cycle and apoptosis genes in addition to c-Myc, Klf4, Oct4 and Sox2 were constructed and used to generate iPSC. The iPSC were extensively characterized by immunohistochemical staining and flow cytometry.

Results: Human mesenchymal stem cells can be efficiently transduced and reprogrammed into iPSC using high-titer lentiviral vectors encoding the four known transcription factors. The addition of siRNA suppressing p53 and cell cycle and survival genes including telomerase and BclXL significantly increased the efficiency and rate of iPSC generation. Human iPSC colonies were formed within a week after lentiviral gene transfer.

Conclusions: The protocol for iPSC generation has been improved with high titer lentiviral vectors encoding additional immortalization cellular factors regulating cell cycle progression, senescence and apoptosis. Deletion of the integrated lentiviral genomes using Cre-loxP recombination could increase the safety profile of the reprogrammed iPSC.

more at http://allelebiotech.com/blogs/2009/12/ascb-abstract-increased-rate-of-reprogramming-of-induced-pluripotent-stem-cells-using-high-titer-lentiviral-vectors-encoding-multiple-cell-growth-and-survival-regulatory-genes/

Construction of An Image Library

The American Society for Cell Biology (ASCB) is “pleased to announce the receipt of a U.S. National Institutes of Health Grand Opportunities (GO) grant to build The Cell: An Image Library. The ASCB will be hiring eight cell biologists or microscopists, each at 25% time,” The job description includes, according to an email job posting, “selecting exemplary images and videos and providing metadata for short tags or descriptions as well as longer annotations including technical details crucial for image interpretations. Annotators will select related key words and note biological source, context, item type, etc., in accordance with set guidelines. Annotators will upload images and videos to the Society’s new image library for research and education.” The grant is in the million dollar range.

The need for creating an extensive image library is deservingly recognized by this “GO” grant from the stimulus program awarded to the NIH by the federal government. The difficult part will be to maintain such an image center once the grant runs out. Will it be kept up-to-date and relevant, or left to collect dust on the old images? We wish that the program would be a great success and that the NIH money well spent.

Allele Biotech has applied to the same round of NIH grants with a related proposal that, rather than cell images in general, focuses more on cell differentiation/dedifferentiation through the use of iPS cells. Title: Foundation for “Subcellular Structureome” as Stem Cell Differentiation Parameters. Summary: The key question to be addressed is how to characterize differentiating stem cells along different lineages definitively and continuously, without disrupting or disturbing the differentiating cells. The broad and long-term goals are to find ways of describing stem cell differentiation in more detailed steps, thereby providing methods to predict and direct cell fate commitment.

Aim 1 Create a panel of cells that can be reprogrammed into induced pluripotent stem cells (iPSCs) with fluorescent protein (FP) fusion markers for each organelle

.Human fibroblasts and keratinocytes will be selected from a large collection of primary human cells, based on their ease to grow and transfect, number of potential cell passages, and potentials for reprogramming with induction reagents. A group of 24 subcellular localization polypeptides (LP) and FP fusion protein constructs currently offered by Allele Biotech will be stably transfected into the selected cell.

Aim 2 Characterize the morphological changes of subcellular structures during iPSC differentiation.

Transfected primary cells that stably express subcellular localization marker proteins will be induced with either current retroviral/lentiviral vectors based reprogramming cDNAs, or a non-integrating baculoviral vector under development at Allele Biotech. These cells, 48 lines in total, will be maintained and expanded under stem cell culture conditions, then induced to differentiate into chondracytes or keratinocytes as examples of cell fate. Morphology data will be analyzed and recorded at each known stage and additional “substage” to be defined in the process.

Aim 3 Correlate morphological changes to known molecular properties of each stage and provide a “signature” set of morphological changes for each stage of each lineage

Signature morphological changes, i.e. significantly different shape, location, sub-type, and copies of organelles in a cell compared to its immediate upstream stage, will be correlated to results obtained by standard expression assays at the RNA and protein levels.

Aim 4 Use the morphology parameters to establish more defined stages of cell fate commitments

Data points will be used to create a novel morphology-based cell fate commitment atlas, which will be very helpful in guiding the stem cell and regenerate medicine research at molecular biology, cell biology and physiology levels.

Aim 5 Construct more FP fusions as organelle-specific markers and combine with stage specific gene promoter driven markers

If necessary, we plan to identify more LPs as fusion marker partners after obtaining the initial set of data, and to expand the signature morphology image database. The database can be further complemented with stage-specific gene promoter driven FP images.

Weekly Promotion of Nov 30-Dec 6: 15% off luciferase assay kit ABP-PA-ABLA011 1000 reactions at only $250.00 212.50. Compare it to what you normally pay for firefly luciferase assays and find out how much you are saving.

Reminder: Allele Biotech Spotlight Promo for ASCB Dec 09 Meeting is still on, order by Dec 9th on iPS and FP groups!

New Product of the Week of Nov 30-Dec 6: Allele Biotech’s ProperFold expression vector with fluorescent protein as indicator for proper protein folding, tracking, and purification. pORB-mWasabi+-sIRES-VSVG