假如我是基醫所新生

賴明德成大醫學院生化所

Starting Academic Career

• If you determine to stay in academic field, obtain a Ph.D. degree first.

• The Ph.D. is a passport valid for immigration into almost any academic institution in the world.

• The ’significance” of Ph.D. study.

• Productive

• Happy

• Dedication

Productive

• Effective working: (1) time management (2) experiment management

• Collaborative working: (1) Novel idea (2) Novel technique

Happy

• Be creative: To verify your creative idea by yourself is one of the happiest things.

• Be independent: It always releases the pressure when the experiment progression is controlled by yourself.

Dedication

• Enthusiasm is the most determining factor for success in science career.

• Dedication to science results from enthusiasm and endurance.

What is your preparation for PHD study?

• Have you done any research before you start to invest your money on stock market?

• Have you done any research before you start your marriage?

• What about the Ph.D. study?

PH.D. Process

• Selecting an advisor: whose lab is right for me?

• Research projects

• Classes, Journal Clubs, Lab meetings, and seminars

• Requirements for graduation: qualify exams

• Do I belong here: stress and insecurity.

Dilemma and Choices for PhD Student

• Advisor

• Research topic

• Perform the experiments

• Write the paper or proposal

• What seminars do I need to attend?

• Do I need to become a Lab’s favorite?

指導教授：研究領域• 興趣是研究熱情的起源• 成就是研究熱情的動力

研究領域之判斷• 你可能對某些領域有強烈之喜好或排斥，

但背後形成的原因卻經常是教師所表現的技巧或熱心，而不是你真正的興趣，或是特定主題所具備的重要性。 Enthusiasm is infectious.

• 學生們常以自己以前作的不錯之題目，當成日後研究領域之選擇基準。

有效率或有創意的實驗室？• We may have many more papers publishe

d when we work in a laboratory performing research efficiently.

• We may learn independence and perhaps be more creative in a less-organized laboratory.

• 李遠哲作為一個例子。

選擇指導教授• 指導教授認定是雙向的流程： (1) 選擇合

適的指導教授； (2)Convince 指導教授 you have the potential to become a good scientist.

合適的指導教授• 對指導教授要有部份的崇拜，但非完全的。

• A graduate student should by all means attach himself to a advisor doing work that has aroused enthusiasm, admiration or respect.

• No good will come of merely going wherever a job offers, irrespective of the work in progress.

選擇指導教授• 最近 5-10 年 publication record

• 過去博士班學生發表論文記錄• 平均修業年限• 過去博士班學生目前職業• 學生的風評 ( 實驗室內或其他實驗室 )

如何讓指導教授認定你 / 妳• 未來潛力• 過去記錄

• 未來潛力： (1) 對面談的準備 (read papers and related material) ； (2) 進實驗室之熱忱； (3) 科學論證能力； (4) 眼神及衣著。

• 過去記錄： (1) publication record ； (2) 碩士論文及摘要說明 (your findings, significance, rationale, experimental techniques)

似乎找不到合適的指導教授• 要懂得處富裕• 也要懂得處卑微

• 使徒保羅

• 努力的漸進改善自己的” recognized scientist potential”.

What shall I do research on?

• Any scientist of any age who want to make important discoveries must study important problems.

• It is not enough that a problem should be “interesting” -almost any problem is interesting if it is studied in sufficient depth.

• Repeat again, Study Important Problems in your field.

What problem is important?

• As an example of research work not worth doing, a young zoology graduate who has decided to try to find out why 36% of sea urchin eggs have a tiny black spot on them.

• Scientists considered collectively are remarkably single-minded in their views about what is important and what is not-whether to science generally or to mankind.

How to evaluate the importance of a science problem

• If the importance of a problem can be inferred from the judgement of most other scientists, there are three ways to judge the importance of your study:

(1) Whether your research results can contribute to mankind health or economy.

(2) Whether a similar work can be published in a prestigious journal.

(3) Whether your seminar on this research work attract audiences in scientific community.

Am I too ambitious in studying important problems?

• The conventional wisdom suggest that young scientist sometimes study the problems which are beyond his/her ability and struggle in the “ important” problem.

• However, it is hard to determine what is the limit of your ability.

• One advice: discuss with your mentor and other scientists before you start a new important topic.

If the topic assigned is not very interesting, then…

• Get the job done, and the paper published.

• Invade another important topic.

• Important science problems are usually solved by collaboration.

Psychology for Scientist• It is psychologically most important to get results, even

if they are not original. As they gain experience, scientist look back upon their own beginnings in research and wonder how they had the temerity to embark on it, considering how thoroughly ignorant and ill-equipped they are. That’s the self-confidence brings them through all these days.

• The final tip: Getting results, even by repeating another’s work, brings with it a great accession of self-confidence.

What shall I do if I have a brilliant idea?

• Is it indeed a creative novel idea?

• Can the experiment be carried out in this laboratory?

• Consult with your advisor!

• Act now!

What to expect the first week

• You will be assigned a lab bench (or part of it) and office space.

• You will be assigned storage space in the refrigerator, -20 freezer, and -70 freezer.

• The lab head, or the person responsible for you, will discuss the experiment you will work on.

• You need to know the biosafety procedure.• You will get keys for the lab.

What to do the first week

• Do an experiment!• Set up your bench.• Introduce yourself to everyone.• Take notes on everything.• Ask! You do not want to bother anyone, especially

needlessly. But it is better to ask experimental procedure than to waste money and time.

• Things you need to find out: chemicals, computer usage, first-aid, glassware, library, lab coats, lab meeting, lab notebooks, ordering supplies, photocopy machine, telephone.

Not to do the first week

• Do not constantly mention, when being shown something, that “we did not do it this way in my class/my other lab.

• Do not read a newspaper or a novel, or play computer games in the lab.

• Do not ask or complain about money or salaries.• Do not use the telephone or photocopy machine

excessively for personal reasons. • Do not suggest that you are working in the lab

for any other reason than love of research.

Survival through common sense and courtesy

• The inconsiderate lab researcher is always a problem to many laboratories.

• If you are going to read anything in this presentation, read this section!

Survival rules: attitude

• Assume nothing. You should not assume someone will immediately stop his experiment to help you whenever you need it.

• Write down everything when someone is giving you instructions.

• Make appointments or request time with people.• Do not discuss fellow lab member’s result with

people not in the lab.

Survival rules: courtesy at the bench

• Never use reagents or buffers without permission.

• If a common reagent is low or run out, order more. Do not put empty bottle back.

• Do not ignore a broken piece of equipment or an equipment alarm.

• Put everything public-use back to original location after use.

• Clean up immediately after each part of an experiment.

• If you do something wrong, confess!

Communication tips

• Forget the still-cherished image of the scientist struggling alone in the lab at midnight, shunning the world. For sure, there will be plenty of late hours with you alone with your tubes, but it is not possible to practice modern science in a vacuum.

• As many discoveries and connections are made in conversations as in biochemical assays, and you must be open to contact with other scientists to make most of their-and your own-research.

Getting along in the Lab (I)

• There are five common problems in the lab: (1) collaborations and credit (2) confrontations (3) favorites (4) deadlines (5) gossips.

• Collaborations: Although most collaborations are worked out before the experiments really begin, they can move in an unplanned directions. The usual problem is that the importance of the individual experiment has changed, and the assumed first-author will be relegated to another person on the credit list. Ask the Principle Investigator to mediate all disputes.

Getting along in the Lab (II)

• Confrontations: Most lab confrontations involve a lab member angry because another member broke a piece of equipment and did not deal with it, did not do the assigned job, used up reagents without ordering more, or used a “private” reagent or equipment without permission. If you are the offended party, deal only with the immediate issue and do not make personal remarks. If you are the guilty party, confess and deal with the problem as soon as possible.

• Another class of confrontation deals with project infringement. It is usually necessary to ask the P.I. to mediate and fix it.

Getting along in the Lab (III)

• Favorites: In the lab, there always seems to be some one who has the P.I.’s attention and admiration. Look carefully, Is it deserved? Maybe you can learn something. And if it is not deserved, mind your own business and learn not let it bother you. Just concentrate on your experiment. The favorite could be you! If so , do not abuse the situation, and do not let it get to your head. You might be out of favor tomorrow.

Getting along in the Lab (IV)

• Deadlines: Follow deadlines, your own and others, religiously. It keeps you organized, and it helps everyone to whom you have made a commitment. Try to set a deadline when someone asks you to do something.

• Gossips: People do talk about people. But be careful. The intimate atmosphere of the lab demands a high level of respect and consideration, even for people you may not like.

How much reading do I need?

About seminar

• Choose carefully the seminars you attend. Do not go to seminars you know you won’t listen to.

• Unless there is a political reason that your body had to be at the seminar, bring your mind also: you do not learn anything by merely show up.

How to attend a seminar

• Listen actively: anticipate where the speaker is going with the data. What is next data?

• Review and summarizing what is being said.• Sorting out evidence and facts from statements

unsupported by evidence. Make judgments, but remain open-minded.

• Asking questions. In an informal seminar, you can ask the questions as they occur to you, but you should wait until the end of a formal meeting to ask most questions.

How to analyze data (I)

• Did the experiment contain “treatment control” and “experimental control”. No controls, no conclusion.

• What conclusion can be reached from this data.• Would the conclusion be changed if some data

are altered. examine the logic in deducing the data.

• What is unanswered? And design next experiment.

How to analyze data (II)

• Some suggestions for the analysis of data:

(1) When data is positive, examine negative control first.(2) When data is negative, examine positive control first.(3) The conclusion should be weaken when certain part of

data was deleted. (4) The conclusion will change when certain data is altered.(5) The “result” of an experiment is never the totality of obs

ervables; the result of an experiment is almost always the difference between at least two sets of observables.

How to analyze data (III)

• Correlation or cause-effect: (1) EGF --> cell proliferation, (2) EGF--> c-fos gene activation ==> correlation. (1) Activation of c-fos --> cell proliferation, (2) Inhibition of c-fos activation by antisense or dominant-negative mutant block EGF-induced cell proliferation ==> cause-effect.

• Biological and chemical observation: (1) UV can induce mutations, (2) the major photoproducts observed in the cell is pyrimidine dimer ==> pyrimidine dimer leads to mutation ?

Do I need to learn many techniques?

• Learn new techniques only when you need it.

How many papers do I need to read?

• Do not read the paper superficially, you need to grasp the essential concept. One example is that knocking out 4 genes can render a normal cells into a stem cells.

• Do not spend too much time in reading a paper unrelated to your current research interests.

• Read the story of discovery.

Harald zur HausenThe Nobel Prize in Physiology or Medicine

2008

New Discovery is usually unwelcomed

• Yes, yes indeed. So, if we may turn to the discovery. When you first suggested that human papilloma virus caused cervical cancer, the prevailing view was that it was herpes simplex virus.

• [HzH] Yes.• [AS] So how was your suggestion at first received?• [HzH] Well it was not very welcome, let's say it this way. Be

cause in the early phase indeed, quite a number of scientists believed that herpes simplex type 2 would do it. There were a number of reports which seemed to confirm that there are increased antibody titres to herpes simplex type 2 in cervical cancer patients. And I reported it for the first time in public in 1974 at a meeting in Key Biscayne in Florida, when there was a meeting which was specifically scheduled for herpes simplex type 2 in cervical cancer. And at that time I reported our negative results in trying to find herpes simplex type 2 DNA in cervical cancer cells. And at the same time I stated that it would be very worthwhile to look rather into genital papilloma viruses, because the reasoning for me was that I had surveyed the literature and found a number of anecdotal reports on malignant conversion of genital warts.

Initial experiments did not support the concept

• HzH] Well, we could demonstrate that there exists a plurality of papilloma virus types. So there was not, as initially assumed, only one single papilloma virus type, but a multitude of different types, which was also confirmed by the group of Gérard Orth in Paris. And, but we concentrated ourselves on trying to isolate the viruses from genital warts because initially we suspected that that virus may be responsible for cervical cancer development. In 1979/1980 we could finally – it was mainly in collaboration with my co-worker Lutz Gissmann – that we could identify HPV6, and Ethel-Michele de Villiers was cloning it subsequently, with Gissmann. And subsequently we could identify another one – that was HPV11, which was also in genital warts and laryngeal papillomatosis. In fact, in the subsequent period, we were initially disappointed not to find HPV6 and 11 in cervical cancer, without one rare exception. But, the probes helped us to identify HPV16 and 18 subsequently. In 1983 and 1984 this was published, by two of my students, Mattias Dürst and Michael Boshart. And, so that was in a way for us a breakthrough because at that time period, of course, interest was awakening very substantially, and quite a large number of groups were collaborating there, and starting to work on papilloma viruses, and ask us for the probes, so we dispersed them very freely throughout the world.

Françoise Barré-SinoussiThe Nobel Prize in Physiology or Medicin

e 2008

Hypothesis must have rationale• [AS] And what gave you the idea of looking for a retrovirus in the first p

lace; the original hypothesis?• [FB-S] First thing is because most of the virus family were already expl

ored. For example, some scientists thought that it could be viruses related to hepatitis virus, or to cytomegalovirus, and so on. The – almost the only family that was not explored was retrovirus. Secondly, as I mentioned before, the only known retrovirus at that time was HTLV-1, and it was reported that HTLV could infect T-lymphocytes. And, of course in this disease, in AIDS, we knew that T-lymphocytes were affected in the disease, characterized by immune deficiency in the patients. Thirdly, I will say that we knew also from the literature that cats, when they are infected by a retrovirus, called feline leukaemia virus, most of the cats are dying from immune deficiency before dying of leukaemia. So, it was several line of arguments telling us that we should look for retrovirus really. So the reason, since we were working on mammalian retroviruses at Pasteur at that time, we said, okay, look, we have all the reagents, we have the technique, and so let's try.

The problem should be important

• For me it's important that the announcement was made at the moment that I was in a developing country because I've been working with developing countries since the mid-1980s and it's really working with those countries that gives me another view, or another way, of orienting my research after the discovery of HIV/AIDS, of the HIV virus. It's important to really know what's going on in those countries strongly affected by these kinds of disease.

Luc MontagnierThe Nobel Prize in Physiology or

Medicine 2008

Medical Importance• AS] Yes, and what is the most urgent research need, do you think, in AIDS

nowadays?• [LM] Well, the important research should be done on

completing the treatments which are given to patients, for life. And I know that in Africa it's not possible to give a treatment for life. Because we give up before. So the idea is to buy a treatment which, like for tuberculosis, could be given for a short period of time – 6 to 9 months – and then stopped. And then vaccinate the people by a combination of viral protein, which I'm working on, so that the immune system of the host, of the person which is infected, will defend himself. Nature has shown us a few percent of people which are in this stage. They are infected, they are not sick. So the idea is to make most of these people, infected people, never sick, for life. So I think this would be very important, but I've tried to quantify all the forms of the virus. So I'm also working on that. There are probably some very small forms of the virus which escape [inaudible] and the immune system. So we have to identify those foremost. I think the AIDS epidemic is caused by a very old virus. And probably the virus was in Africa for a long period of time, as shown in some recent papers.

Observation is the key element

• AS] Yes. So my last question. If you remember the day on which you first saw reverse transcriptase activity in the cultured cells from patients. Did you have any idea on that day in 1983 of the size of epidemic that you might be looking at?

• [LM] Well not in the beginning of course, no. I was actually working on a possible viral cause of breast cancer. I'm still interested in cancer virus so I appreciate that the Nobel Committee also has awarded the Prize to Harald zur Hausen who has worked for a long time on this. But, when I realised that the virus could be the cause of AIDS and was present not only in gay men in France and the United States, and haemophiliacs, but also in African nations – so this was in September 1983 – I realised it could be big, okay?

Roger Y. TsienThe Nobel Prize in Chemistry 200

8

We try to build molecules to solve problems.

研究興趣 is the key to success

• AS] When did you actually realise you were a scientist?• [RT] I guess I realised I was most interested in being a s

cientist in the ... Oh I guess it would have been junior high school, say, probably the age of 9 or 10.

• [AS] That's pretty young, that's pretty young.• [RT] Well I, yeah, I guess I think ... I'm trying to reme

mber when I started playing around with chemistry experiments at home. It was probably around then. Much to the horror of ... Well, a mixture of fear in my parents and a little bit of encouragement.

Insecurity and Stress (I)

• Other students seem to know everything, but I….

• What made me feel better was remembering what I overheard a visiting professor say” Postdocs do not know very much-they are still students” (Ph.D. Cell Biology, Baylor College of Medicine)

• Solution: If you are not so smart, why are you here?

Insecurity and Stress (II)

• “I almost never talked in Journal club. The only time I spoke was when I was required to do so. That was scary; I hated it” (Ph.D. Biological Sciences, University of Texas at Austin)

• Solution I: with time, you will understand the jargon and things will make more sense.

• Solution II: (1) read the paper in advance (2) try to phrase your question before you ask it (3) figure out the type of questions others asked.

Insecurity and Stress (III)

• “There are so many sources of stress that is hard to know where to start. The primary ones were lack of time and perpetual shortage of money. Anxiety over qualify exams were also high on list.” (Ph.D. Zoology, University of California, Los Angeles)

• Solution: Unfortunately, the stresses involved in becoming a Ph.D. candidate cannot be avoided. Joining a student supporting group can be helpful in relieving tension. On the other hand, many students really “take off” when they finally start doing experiments.

• Error can often be fertile, but perfection is always sterile. – A.J.P. Taylor

Success of a Ph.D. degree

• Aim: pursue the beauty of knowledge.

• Sub-Aim: get employed.

Take home advice

• 創造自我價值： make yourself be important to others.

• 創造他人價值： make others be important to you.

References

1. “Advice to a young scientist” by Peter Medawar, 1981.

2. “At the bench, a laboratory navigator” by Kathy Barker, 1998. Cold Spring Harbor Laboratory.

3. “Who wants to be a scientist” by Nancy Rothwell. 中譯本：科學家的第一堂課，陳佳伶譯，早安財經文化有限公司， 2004 。

4. “The Ph.D. Process” by Dale F. Bloom, Jonathan D. Karp, and Nicholas Cohen. 1998, Oxford University Press.