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VOLUME l 16 APRIL 2009l l

Journal of Indian Academy of NeurosciencesOfficial

PUBLISHED QUARTERLY www.neuroscienceacademy.org.in

Annals of Neurosciences

ISSN 0972-7531

A SN VOLUME 16 NUMBER 2 APRIL 2009

Annals of Neurosciences Official Journal of Indian Academy of Neurosciences

Editor-in-Chief

Akshay Anand, Ph.D

Neuroscience Research Lab

Department of Neurology,

Institute Stem Cell Facility

Post Graduate Institute of Medical

Education and Research, Chandigarh

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S Prabhakar, D.M

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P. Joshi, Ph.D

Ante Padjen, M.D, D.Sc

Y. K Gupta, M.D

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Mriganka Sur, F.R.S

Obaid Siddiqi, Ph.D, FNASc

S C Lakhotia, Ph.D, FNASc

Peter Andrews, Ph.D

Keiji Tanaka, Ph.D

Martin Schwab, Ph.D

Hector H Garcia, M.D, Ph.D

Tom Isaacs

K. Satyanarayana, Ph.D

Zhen-Ge Luo, Ph.D

Adhibato Rao, Ph.D

R K Vasishta, M.D

Bimla Nehru, Ph.D

Nibedita Lenka, Ph.D

Rakesh Biswas, M.D

Nusrat Shafiq, D.M

Asok Mukhopadhyay, Ph.D

F. Islam, Ph.D

Paramjeet Singh, M.D

Almaz Aldashev, Ph.D

Stefan Gluck, M.D, Ph.D

G. U. Gurudatta, Ph.D

R.A.P. Alcala, Ph.D

Nihar Ranjan, Ph.D

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IP Consultants

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Michael Coblenz

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Rakesh Shukla, D.M

Founding Editor

J.N. Sinha, Ph.D

Editorial

New Focus

Commentary

Research Article

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Classics

Comprehensive Review

Mini Review

Report

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46

48

49

54

56

57

62

72

75

the long and short of it

Akshay Anand

merit, seniority and science

Suptendra Nath Sarabadhikari

asking the right question – the ignored key of medical research

Nusrat Shafiq

A modified, rapid fluorometric method for the quantification of mitochondrial calcium

Amit S. Korde and William F. Maragos

Cells previously identified as retinal stem cells are pigmented ciliary epithelial cellsPNAS 2009 ; 106 : 6685-6690

Chandrika Abburi

Mind – Body dichotomy

Vinod Srivastava

Discourse on the method of rightly conducting the reason and seeking truth in the sciences (Contd. from Previous Issue)

Rene Descartes

Age related macular degeneration - advances and trends

Neel Kamal Sharma, Sudesh Prabhakar and Akshay Anand

Ovarian hormones and the brain signals

B.W. Gawali, P.B. Rokade, G.B. Janvale and S.C. Mehrotra

3rd Canadian IBRO school of neuroscience : neurodegeneration and regeneration

Akshay Anand

On Page 80, MAP2 immunostaining of neurons

On Page 79Book Review

On Page 82, ANS announces new editorial team

ANNALS OF NEUROSCIENCES

VOLUME 16 NUMBER 2 APRIL 2009

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Case Report

Book Review

Molecular Shots

People and Views

77

the new team

Dissemination of tubercular bacilli by shunt placement

Sandeep Mohindra, Rahul Gupta and Rajesh Chhabra

Animal Cell Technology by Asok Mukhopadhyay

S.N. Mukhopadhyay

Immunostaining pictures

Gururaj Joshi and Jeffrey A Johnson

Nibedita Lenka, Hoon-Ki Sung and Paria Mohseni

Letters to Editor

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80

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ANNALS OF NEUROSCIENCES

VOLUME 16 NUMBER 2 APRIL 2009

On Page 62, AMD Review

On Page 77, Unique case of TBM

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ANNALS OF NEUROSCIENCES VOLUME 16 NUMBER 2 APRIL 2009www.neuroscienceacademy.org.in

A SNEDITORIAL

This editorial is being written at a time when considerable intellectual portfolios and are The new law provides opportunities to Indian (neuro)science is witnessing a major ready to seek new frontiers in the field of aggressively establish for itself a corpus of policy shift in redefining the Indian research. technology and national wealth creation individuals which can generate wealth for it.

2Today we are confronted with a much endeavours . Many view this new regulation The new provision allowing mobility of awaited avatar of Bayh-Dole Act allowing for as serving to create a distinct reward system faculty to and fro from industry, can the transfer of exclusive control over many for over performers over average workers accelerate seamless transfer of knowedge government funded invent ions to and may have potential to reverse brain from one organization to other and industry. universities, Institutions and businesses for drain. This could serve as a good tool for This regulation can now provide competitive the purpose of further development and growing Institutions that are apparently not edge to an Institute which is bestowed with commercialization. This bold initiative able to support their own thought leaders faculty with special skills and entrepreneurial communicated by Ministry of Science and for want of stringent service conduct rules. drive than those Institutions that don't. Technology seeks to transcend mere The leadership displayed by IIT entrepreneur- Therefore, an early composition of task licensing of inventions to third parties to ship schemes is laudable for others to forces to oversee the establishment of such strengthening scientists' claim of holding emulate. Nusrat shafiq argues in her offices will be the key to consolidation of an equity in scientific enterprises and creating commentary in this issue why there is early lead in the field that will now become spin offs, exempting them from the lackadaisical approach in asking good intensely competitive and attractive. Such a draconian cluthches of CCS conduct rules. research questions, makes one think why the set up is bound to stimulate product (or This recent Govt order is meant to ensure the Indians who are settled abroad perform patient) oriented research in anticipation of continued involvement of researchers in better than those who live in this country. profits. Many see a dream opportunity for translating the inventions or innovations, at Perhaps the right individuals with right medical Institutes to translate this regulation any stage of development, through questions can now expect to be rewarded into establishment of Business Development investment of their own money. This has from taking risks of asking the right office comprising people from Knowledge been elaborated in the draft that sets the questions and creating spin offs. Supten, on management (such as IP management), guidelines about creation of scientist based the other hand, argues in his commentary informatics, quality control and as venture companies thereby permitting researchers to why number of years in service should be a capitalists.play a defining role in commercialization of necessary replacement for merit in science. knowledge products while being in Young entrepreneurs in scientific journalism Like developed nations, where excellence is professional employment. The entire draft too have a new opportunity to combine their the hallmark of career(read national) has been published at www.dsir.gov.in for enthusiasm, ideas and productivity with advancement, India too needs to experiment the information of its readers. The new law is advisory role of accomplished scientists so with its policies such that many Gokhales are applicable to all Institutes falling under that a new order of thought leadership is given leadership roles of research Ministry of Health and Family welfare, created in running journals. Journalism is organizations, journals and academies. It is besides other scientific establishments, likely to gain because many may opt to start therefore not difficult for India to regain its providing a unique opportunity for medical new journals as a spin off. New ideas of global leadership in the field of Science and institutes to mobilize its intellectual enhancing visibility to journals will include Education. It would, therefore, be very resources to generate wealth, an effort never experiments with its design, content, interesting to see how the new regulation is envisioned earlier. Some feel that this is an enhanced investments, new breed of shaped in the context of authority (read opportunity to initiate incubation centres in 3publishers, venues and leadership roles . seniority) patterns identified in Supten's their campuses and mobilise its human commentary. This will eventually lead to new dimension of resources to industry and recall them back neuroscience publications in the country. Given the impact stem cell research is making later to establish their own business

Some will question publication ethics, others in transforming regenerative medicine, this development offices for generation of the national funding mechanisms and new law can enable researchers to create research funds. Both these measures have policies, some the value of research audit, practical solutions by meeting social been sanctioned in the government order some the scientific dogmas and yet others challenges and to directly participate in with the expressed purpose of supporting the science leaders themselves.1 creating and sustaining competitive, self internal fund generation projects . Such new

financing and low cost patient care industry wealth can be harnessed for rationalization Akshay Anandfor decades to come. This can be of patient care costs as well as funding more

Referencesconceptualized by mobilising investment of R&D projects within the Institute. The 1. Anand A, Science entrepreneurship: several business houses who might want to potential of this approach in transforming

Challenges and Opportunities in India, set up their centres within the institutes, regional economies (and that of the Journal of Public Administration and particularly medical organisations, and fund Institute) by creation of new companies is Policy Research 2009;1(1): 1-3

research in return for commercialisable borne out of the examples set by MIT, IIT, IIM, 2. Das P, Economic liberalisation and R&D and products. At a time when organisations like Stanford and University of Cambridge, many

innovation responses of Indian public CSIR, IIT and IISc have devised mechanisms to of which are based on biomedical inventions. and private sector industries. Int J Manag open incubation centres, medical institutes By promoting science and engineering based Decision Making 2004;5(1): 76-92

enterprises it has not only opened vistas for in the country have a unique opportunity to 3. Anand A, Journal needs aggressive policy, Institutes to impact society directly but also make impact in the field of biomedical Editorial Annals of Neurosciences paved way for those who possess innovation and not be left behind in the race. 2009;16(1): 1.


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ANNALS OF NEUROSCIENCES VOLUME 16 NUMBER 2 APRIL 2009 www.neuroscienceacademy.org.in

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3Meritocracy, in practice, means the Vardas observes that apart from the ? c a n e x p e c t a d v a n c e m e n t selection of the “best and the brightest” opportunities, either formal or evident decline of the medical education for positions of power and authority, and informal, within her institution, and research as a whole, it is particularly is usually supposed to be independent of apart from other criteria.disquieting to see the appointment of age. The antonyms for meritocracy are administrative leaders within the Greek

Unfortunately, in India, the above criteria nepotism, favoritism, preferential health system, or even sometimes of are not defined while promoting or 1treatment and privilege . The opportu- university professors, with barely a nod to appointing academic administrators or nities for leading medical institutions in meritocracy. Often, the choices reflect policymakers. Another example of similar India are usually given to individuals based political friendships, cliques and blocs, initiative is the Academic Administrator, on considerations other than their which are usually players in a variety of 5Clinician Educator (AACE) Track that was personal merit. In India, “age” is usually power games. While the relative developed as part of a movement towards s y n o n y m o u s w i t h “ w i s d o m ” . depreciation of undergraduate studies promoting future academic leaders in Unfortunately, while experience and and degrees is alarming, the failure to clinical care, continuing education, and performance are important, nothing can appoint the most competent individuals administration. As part of the application really ensure that age will necessarily to leadership positions within the system process, the candidate must propose an endow anyone with the skills required of can have potentially devastating medium academic project (e.g., Quality Assurance becoming more productive or efficient to and long-term consequences. To further activity, data-mining investigation, propel the country forward in a fast complicate matters, the mediocre publication of clinical findings, or moving field of modern medical research. managers demonstrate day after day their educational initiative/course/seminar). I am citing two articles from Iran and inability to organize models for develop- Such academic leadership programs are Greece to show that meritocracy is often ment, to adapt to modern competitive also offered globally through the the least (and last) important factor for requirements or to inspire their juniors. Foundation for Advancement of consideration of appointment for

International Medical Education and The Hedwig van Ameringen Executive academic and administrative heads for 6Research (FAIMER) that was incorporated 2,3 Leadership in Academic Medicine® medical education and research

as a nonprofit foundation of ECFMG (ELAM®) Program for Women is an

Arguably, many academic leaders who (Educational Commission for Foreign example of an in-depth program focused

lack the required merits or appropriate Medical Graduates) in September 2000. on preparing women for senior faculty

qualifications, to excel and tend to be FAIMER Regional Institutes are modeled positions as well as leadership positions

conservative which needs to change if the on the FAIMER Institute curriculum. They where they have effectively initiated

country has to sustain momentum in include residential sessions, as well as positive change in the United States. This 2medical research. Bikmoradi et al report distance learning sessions. Each program is supported and hosted by

that the criteria supporting academic participant is also required to propose and 4Drexel University which believes that an leadership are usually relegated to implement an education innovation

ideal candidate for the program should be politicization, informal groups, and project that is supported by the home one whoexternal forces in Iran. Perhaps these institution. To date there are five Regional

factors pose limitations among academic Institutes, three in India (at GS Medical ? has attained at least the rank of leaders and create unpleasant experiences College, Mumbai; CMC, Ludhiana and PSG Associate Professor and has about utilizing their authorities. The IMS & R, Coimbatore), one in Brazil, and achieved significant administrative advancement of field is seriously the Southern Africa Regional Institute, experience in personnel and budget compromised when many young brains which began in February 2008. Therefore, matters, preferably both (e.g., become hesistant to challenge a decision it is high time that we overcome the inertia Department Chair, Division or on distribution of funds or question of past and make use of such innovative Section Chief)established research policies and scientific programs to change the rules used to

? expresses a clear desire for attaining paradigms for want of opportunities to select key individuals of our research a leadership positioninfluence decision making. To continue to infrastructure to ensure continuing

develop life saving technology, save lives advances in this important area. We ? embraces strategic risk-taking as of millions of countrymen who suffer from cannot rely on foreign entities to fulfill this

part of her career pathdreaded diseases, the selection of role, as these entities shall compete with

? realistically assesses her leadership individuals, irrespective of age, is must for India, not work in conjunction with us, to accelerating the development of medical opportunities, both internal and attain new results. India provides a wealth sciences. external of opportunity in the arena of medical

merit, seniority and science

NEW FOCUS

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A SNNEW FOCUS

2. Bikmoradi A, Brommels M, Shoghli A, Sohrabi research as well as a huge patient base to needed to arrive at this enviable position. Z, Masiello I. Requirements for effective explore innovative approaches. India We can attain the facilities; the personnel, academic leadership in Iran: a nominal

offers intelligent investigators many of the funds if we sustain the desire to group technique exercise. http://www. whom have set the example for progress achieve this goal independently, but only biomedcentral.com/1472-6920/8/24

BMC Med Educ. 2008 ; 8:24working in other countries. Scientists from if we face brutal realities that change the US and other countries have found in necessitates. Let us select our adminis- 3. Vardas PE. The ant, the grasshopper, and

meritocracy: where has Aesop's fable India an environment that fosters trators after we assess their vision, their g o n e w r o n g ? h t t p : / / w w w . excellence in identification of therapeutic competence, their ambition rather than hellenicjcardiol.org/ archive/ full_text/

advance, and India has become in 6 short their age. 2007/4/2007_4_251.pdf Hellenic J years a major presence in the world stage Cardiol. 2007 ; 48(4): 251

Suptendra Nath Sarbadhikari,of research innovation. Let us not relegate 4. Drexel University College of Medicine, Editor Journal of Indian Health and confine our future advances to the C a n d i d a t e S e l e c t i o n C r i t e r i a : Informatics, http://www.drexelmed.edu/Home/Otherinput of investigators from other nations

Programs/ExecutiveLeadershipinAcademDepartment of Biomedical Informaticsfunded by industries and governmental icMedicine/ApplicationProcess/Selection

entities that have an admirable agenda, PSG Institute of Medical Sciences and Criteria.aspx (August 12, 2009)Research, Coimbatore 641 004, Indiabut not an agenda that is directed at

5. Department of Psychiatry, University of advance in our country; at minimizing the Email: [email protected] Pittsburgh School of Medicine, Clinical diseases and suffering our country's Educator and Academic Administrator Co-op. Editor : Denis English, Ph.D

Training during Residency - AACE Track: population experiences; at propelling Senior Editorhttp://www.wpic.pitt.edu/education/resiIndia functional and intellectual parity dency_tra in ing/AACE_TRACK.htm

with the recognized leading nations in (August 12, 2009)Referencesmedical research. We are almost there. We

6. FAIMER Regional Initiatives: http://www. 1. Saleh JH, On Values and a caring Meritocracy possess the initiative, innate intelligence faimer.org/ education/regional.html for MIT. http://web.mit.edu/ fnl/volume/ and imagination, population and freedom 182/saleh.htm (August 11, 2009) (August 12, 2009)

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A SNCOMMENTARY

asking the right question – the ignored key of medical research2Medical science has progressed by leaps ago. ceutical industry devoting much more on

and bounds in the last century. The credit research and development of novel Many question if the funds are reaching of this progress rests with research done molecules. Some of them have invested in the right projects. There is indeed a review both in basic and clinical fields. A quick infrastructure for research and develop-process wherein the research projects are review of the published literature, ment. However, we lack a vision that could sent to reviewers for their comments. however, shows an abysmal contribution chase the dream of addressing the unmet Either the best ideas are not getting from our country. This doesn't imply that medical application. In the end, the funded or there is a dearth of worthy Indian names do not figure in the pharmaceutical industry has either kept project proposals. A good project should published medical literature. In fact, most aloft our ability to produce generics or be determined mainly by the question that of the path breaking studies have names have progressed to form mergers.is being asked or the problem that is being which are obviously of Indian subconti- addressed. Novelty is often missing in After the Jaipur foot, our contribution to nent. This is indeed heartening. However, majority of the projects. It is rare to find a an innovation in the field of medical we cannot ignore the fact that these research project that addresses an instrumentation and technology has research contributions have been carried important unanswered problem which become conspicuous by its absence. Some out in USA and European Union, often may be related to clinical practice or initiatives have certainly been taken in this carried out by post doctoral fellows who understanding of a prevalent disease.

respect. Funding agencies are creating are imported by the dozens every year. In How many have really been equipped with public private interface where the need the past two decades, several scientists tools to assess the worthiness of the and technology development can be who were t r a ined i n r e sea r ch project from such perspectives. It is also brought together. However, again we fall methodologies abroad came back to the difficult to understand, why our short of ideas. Intracoronary stents, heart country. They brought with them new outpatient clinics, which are thronged by valves, magnetic resonance imaging, techniques and established some good patients of all kinds, do not lead to none have been conceived in our hospitals laboratories across the country. Yet, the description of a new syndromes. We often

research climate did not change much. and in our laboratories but invariably have people lamenting about lack of time However, there was one transformation- imported. We tend to devote more time in to do research. While that may be true, it is increased sensitisation about research in perfecting angioplasty technique rather also true that such a diversity of cases the medical Institutions. The need for than being the first one to ask, 'can some opens up a person to make new obser-research has been realized. Yet, we do not mechanical device be developed to open vations and ask many unanswered contribute, in tangible ways, to the up the blocked vessel?' questions. Astute observations and asking multitude of novel findings which appear resolved medical questions are two We need to guide our research by good in the indexed journals. Where does the entities which are resource independent. questions that influence our practice of problem lie? A quick look at the website of Once this process starts then first step of medicine and immediate society. Good Duke University School of Medicine bedside to bench research can be research design and methodology are reveals a list of Duke's first which range achieved. A research question which has indeed needed. Some mechanism will from simple and yet important its origin in local problems has a greater need to be set up wherein the person who innovations to some more complicated potential for bringing paradigm shifts.

1 has these qualities of making novel and equally important findings. Perhaps the orientation of research has to observations and asking good questions is It is convenient to blame the lack of be such that it transforms the bedside encouraged to play a leadership role appropriate funds for the present state of experience into a benchtop exploration.

backed with a team who can design and affairs. However, going through the list of This highlights the need to enhance

conduct good basic and clinical research.granted projects and the volume of commitment towards meaningful and

Nusrat Shafiqfunding for many projects, it lead us to applied research. One way to begin is to Department of Pharmacology, believe otherwise. Others argue that expand something that is developed by Post Graduate Institute of Medical Infrastructure and facilities for conducting thought leaders and then extend upon it. Education and Research, Chandigarhresearch may be limited in many After J receptors, novel findings have

Institutes, however, there are others who taken a backseat. Indeed a lot of E-mail : [email protected] well equipped laboratories and yet unanswered questions have been References:lack ability to cause major paradigm shift addressed. However, there is still a gamut

1. http://medschool.duke.edu/ modules/ in medical research. It is sad to note that a of clinical queries which need to be taken som_rt/index.php?id=1drift in the thinking process has occurred care of. Few years ago when product

2. Arunachalam S. How relevant is medical yet no major productive shifts are patent was introduced in India, a lot of research done in India?- A study based on noticeable as pointed out nearly a decade people were upbeat about pharma- Medline. Curr Sci 1997; 72: 912-22

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A SNRES ARTICLE

A modified, rapid fluorometric method for the quantification of

mitochondrial calcium1 2,3,4,5Amit S. Korde and William F. Maragos

1 , 2 3 4Center for Cardiovascular sciences, Albany Medical College, Albany NY USA Depts. of Neurology , Anatomy and Neurobiology , Graduate Center for Toxicology 5and Sanders Brown Center on Aging , University of Kentucky, Lexington, KY, USA.

ABSTRACT

Calcium is an important cellular mediator in numerous physiological processes. Under pathological conditions, 2+large influxes of Ca into mitochondria may occur, leading to activation of “cell-death” pathways. Although

2+ 2+there are several methods by which to monitor mitochondrial Ca [Ca ] in vitro, current methods for m2+quantifying [Ca ] in the living animal are limited. Herein, we describe a rapid and simple method by which to m

2+ 2+quantify the levels of total [Ca ] in situ. The method relies on the presence of inhibitors of Ca influx and efflux m2+to prevent movement of Ca during mitochondrial isolation followed by the lysing of the organelles to liberate

2+the cation. Total [Ca ] is quantified using the fluorescent probe Calcium Green 5-N. The method is sensitive to in m2+vivo perturbations, as demonstrated by decreases in [Ca ] following administration of the uncoupling agent 2, m

4-dinitrophenol.

KEY WORDS

Mitrochondrial calciumCalcium green 5NFluorometryLocking buffer

Corresponding Author

Amit S KordePhone (518) 262-6477Fax (518) 262-8102email: [email protected]

Introduction information of subcellular calcium levels in the whole animal. 2+ Recently, a method was devised that involves the isolation of Calcium (Ca ) has been recently recognized as a critical factor in

1 mitochondria in a buffer that contains the cation chelator EDTA the mitochondrial control of cell death . It is now well 2+

2+ to minimize free Ca , ruthenium red to inhibit the mitochondrial documented that when cytoplasmic Ca is increased, 2+ 2 2+ uniporter and voltage-dependent anion channel (VDAC) and mitochondria act as sinks for Ca . Indeed, mitochondrial Ca

+ 2+ + 2+2+ lacks Na to prevent extrusion of Ca via the Na / Ca concentrations ([Ca ] ) can increase from a physiological m 8antiporter . The mitochondria are then ruptured using Triton X-concentration of approximately 100 nM or less to greater than

100 and liberated calcium quantified using the fluorescent 5µM under pathological conditions. Although physiological +probe fura-2K . A potential limitation of this method is the 2+ increases in cytoplasmic Ca are readily buffered by the presence of EDTA in the isolation buffer which might shift the mitochondria and then rapidly redistributed, abnormally high

2+ equilibrium of Ca from inside the mitochondrial matrix to the 2+levels of [Ca ] can be deleterious and may initiate a cascade of m + 2+buffer via efflux through the H /Ca antiporter and/or the 1 2+events leading to either necrotic or apoptotic cell death . Ca - +permeability transition pore (PTP). In addition, because fura-2K mediated cell death may represent a final common pathway in a is a ratiometric dye, calcium concentrations must be derived number of chronic and acute neurodegenerative conditions using a standard curve that requires the determination of the 3including Huntington's disease and stroke . minimum fluorescence using a buffer containing EGTA and

In order to completely understand the molecular events leading maximum fluorescence following the addition of saturating 2+ to cell death in animal models of neurodegeneration, it is concentrations of Ca . In the method described herein, we have

2+necessary to quantify [Ca ] in vivo. Currently, there are several m circumvented these issues by isolating mitochondria in a means with which to accomplish this. One such method relies on “locking buffer” that contains no chelators and includes

2+ 2+the infusion of [Ca ] into the live animal followed by inhibitors of currently described mitochondrial Ca 2+autoradiographic visualization of silver grains using electron channels/pores to prevent either the influx or efflux of Ca

4microscopy . Yet another method utilizes the oxalatepyroanti- during tissue processing. In addition, the use of the non-manate technique to visualize intra-mitochondrial calcium ratiometric fluorescent probe Calcium Green 5-N, allows the

5precipitates using electron microscopy . Although both of these liberated calcium concentration to be extrapolated from an methods provide high resolution, limitations of the former easily generated standard curve.method includes the use of radioactivity and both methods are

Methodslimited by the difficulties associated with quantification and tissue processing for imaging. More current methods rely on the Six to eight week-old (approx 30-35 grams) male C57 mice were

6use of fluorescent probes and visualization using either single or used for all animal studies. All animal use procedures were in 7two-photon confocal microscopy. These semi quantitative accordance with the NIH Guide for the Care and Use of

methods, too, are sensitive and have proven useful in tissue Laboratory Animals and were approved by the Albany Medical culture studies; however they lack sufficient resolution to afford College Institutional Animal Care and Use Committee. CGP

37157 was purchased from Tocris Inc. (Ellisville, MO). Ruthenium mitochondrial uncoupler that can be used to decrease the red, cyclosporin A, Triton X-100, 2,4-dinitrophenol (DNP), DMSO mitochondrial membrane potential (ψ). Since the influx of m

2+ 13,2and 2,4,6-trinitrophenol (TNP) were purchased from Sigma (St. cytoplasmic Ca into the mitochondria is dependent on ψ, m

Louis, MO.). Calcium Green 5-N and BECEF acid (2',7'-bis- we hypothesized that treatment with DNP would result in a 2+(2-carboxyethyl)-5-(and-6-)-carboxyfluorescein) were purchased reduction of [Ca ] . One hour following administration of 5 m

from Molecular Probes (Eugene, OR.). All other chemicals were mg/kg DNP (in 60% DMSO), animals were sacrificed and reagent grade. mitochondria were prepared from forebrain as described above. In our initial studies, mitochondria were prepared from whole In order to control for any non-specific effects of DNP, a control

9forebrain using a modification of the method of Sims . In this group of animals was administered the molar equivalent (6.2 assay, the whole brain was placed in 2 ml of isolation buffer (20 mg/kg in 60% DMSO, i.p.) of the DNP analogue 2,4,6-mM HEPES, 1% BSA, 75 mM sucrose, 215 mM mannitol, pH 7.2) trinitrophenol (TNP). Although TNP can uncouple submito-that contained 1) 16 µM ruthenium red to block the chondrial particles in which the inner mitochondrial membrane

2+ mitochondrial inward transport of Ca via the uniporter and is inside-out, it is impermeable to intact mitochondrial 14extrusion via the sodium-independent antiporter and voltage- membranes and thus would not be expected to alter ψ and, m

2+dependent anion channel (VDAC) 2) 10 µM cyclosporin A to consequently, [Ca ] . Data were analyzed using Student's t-test m2+ prevent loss of Ca via induction of the permeability transition and are expressed as + SEM.2+ and 3) 15 µM CGP-37157 to block outward flux of Ca via the

Resultssodium-dependent antiporter. The concentrations of the inhibitors used to make this “locking buffer” approximate the A standard curve was generated using locking buffer that concentrations that were successfully used in other in vitro contained 50 µg mitochondrial protein to which known

10,11,12 + 2+preparations . Neither Na , which is required for activation concentrations of [Ca ] was added. The corresponding of the sodium-independent antiporter, nor EGTA/EDTA was fluorescence values were linear in the range from 100nM to 200

included in the locking buffer. The final pellet was taken up in M and the equation (y=mx + C) was used to describe the curve the same buffer and 25, 50 or 100 g mitochondrial protein were (Fig. 1, top curve). To make certain that the fluorescent signal

2+ placed in the wells of 96 well plates to which was added 5 µM resulted from the Ca itself, another standard curve was Calcium Green 5-N in a final volume of 100 µl locking buffer. generated in the presence of 100 µM EGTA and resulted, as

2+Total [Ca ] was determined by subtracting the background m expected, in the reduction of the fluorescent signal to levels fluorescence of the intact mitochondrial preparation from the nearing zero (Fig 1, bottom curve). A parallel set of standard fluorescence intensity following solubilization of the organelles curves generated in the presence of either lower (25 µg) or with 100 µM Triton X-100 15 min. Calcium Green 5-N has the higher mitochondrial protein levels resulted in a slope that was advantage over esterified mitochondrial permeable probes in essentially identical to that seen with 50 µg protein and indicates that it is not dependent on the mitochondrial membrane that the amount of mitochondria in the assay mixture does not potential, does not require enzymatic cleavage by esterases and, interfere with the fluorescent signal. once the mitochondria have been solubilized, will measure total

2+[Ca ] (i.e. bound + free). Samples were read in 96 well plates m

using a BIO-teck Synergy HT fluorometer using an excitation wavelength of 485nm and emission wavelength of 532 nm. Tissue concentrations were extrapolated from a standard curve

that was generated using known concentrations of CaCl .2

Protein concentrations were determined using the Bradford protein assay (Pearce, Rockford, IL). All experiments were

2+replicated a minimum of three times and [Ca ] is expressed as m

nmol/mg mitochondrial protein.

Due to the ability of Calcium Green 5-N to fluoresce with changes in pH, we conducted a series of experiments to determine whether solubilization of mitochondria and the release of protons altered the buffer pH. Following preparation of mitochondria in locking buffer, tissue samples were incubated for 15 minutes with 10 µM of the fluorescent pH indicator BECEF before and after addition of 100 µM Triton X-100. Samples were read in 96 well plates with an excitation wavelength of 485 nm

Figure 1. Standard curve (top curve) showing linear relationship and emission wavelength of 532 nm. The pH of the samples was

between Calcium Green 5-N fluorescence signal intensity as a result calculated by extrapolating the fluorescence from a standard

of increasing concentrations of CaCl (top curve). The assay was 2curve that was generated using buffers of known pH and linear carried out in the presence of 50 µg mitochondria in “locking” buffer

in a pH range from 4 to pH 10 (not shown). containing 16 µM ruthenium red, 10 µM cyclosporin A and 15 µM

To determine whether this method could detect changes in CGP-37157. The bottom standard curve demonstrates the effects of 2+[Ca ] in vivo, we treated a group of animals systemically with 100 µM EGTA and indicates that the fluorescence intensity signal m

2+ 2,4-dinitrophenol (DNP) (5 mg/kg in 60% DMSO i.p.). DNP is a was the result of Ca ions.

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A SNRES ARTICLE 50


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2+We next determined the background [Ca ] using various protein contrast, animals treated with DNP, showed an almost 60% 2+concentrations of intact brain mitochondria from normal, reduction in [Ca ] compared to mitochondria from control m

2+ untreated animals. Although the absolute amount of Ca animals (2.81 + 0.148 nmol/mg vs. 6.99 + 0.136 nmol/mg predictably increased with greater amounts of mitochondria, protein; p<0.001). To control for any non-specific effects of DNP, the values were essentially the same when these were a third group of animals was systemically administered the

2+ normalized to total protein. Specifically, background Ca was molar equivalent of the structural analogue TNP. One hour 2+0.37 + 0.003 nmol/mg protein in 25 µg samples, 0.36 + 0.012 following administration of TNP, [Ca ] was 7.06 + 0.142 m

nmol/mg proteins in 50 µg samples and 0.38 + 0.009 nmol/mg nmol/mg protein, which was not significantly different from proteins in 100 µg samples (Fig. 2). Following solubilization of DMSO-treated animals (Fig. 3).mitochondria with Triton X-100, there was again a marked

2+increase in [Ca ] that, when normalized to protein levels were

independent of the amount of mitochondria. In this experiment, 2+(Ca ] was 7.63 + 0.047 nmol/mg mitochondrial protein using m

25 µg samples, 8.06 + 0.097 nmol/mg mitochondrial protein in 50 µg samples, and 7.77 + 0.155 nmol/mg mitochondrial protein in 100 µg tissue samples (Fig. 2). Due of the similar results obtained with different concentrations of protein, we elected to use 50 µg of mitochondrial protein for the remaining experiments. Additionally, in view of the lack of variability of individual standard curves over time, a single standard curve in which 50 µg mitochondria protein was included in the buffer, was generated by averaging 6 curves prepared on sequential days and used in all subsequent experiments. Figure 3. Effect of the mitochondrial uncoupler DNP on in vivo

2+ 2+[Ca ] . The background Ca in buffer containing intact m

mitochondria did not differ between treatment groups. Following 2+solubilization with Triton X-100, Ca concentrations in both DMSO-

2+and TNP-treated groups increased markedly and [Ca ] was not m

different from that seen mitochondria prepared from normal, untreated animals (see Figure 1). In contrast, there is a significant

2+decrease in [Ca ] in mitochondria prepared from an animal treated m

with 5 mg/kg DNP (p < 0.001), consistent with a decrease in the mitochondrial membrane potential.

2+ Figure 2. Effects of increasing protein concentration on Ca levels in intact and Triton X-100 solubilized mitochondria. In the intact mitochondria, there is no decrease in background concentrations of

2+Ca as protein concentrations increase. Following exposure to Triton 2+X-100, there is a marked increase in [Ca ], presumably due to the

2+ release of both bound and free Ca from the mitochondrial matrix. 2+As expected, the concentration of [Ca ] was constant and m

independent of the amount of mitochondria assayed. Figure 4. Effects of in vivo administration of DNP or DMSO on assay

To determine whether this method was sensitive to changes in buffer pH. Animals were administered systemically either 5 mg.kg 2+ Ca mobilization in vivo, we treated a group of animals with DNP or the molar equivalent of DMSO. One hour later, forebrain

2+DNP and another with DMSO and compared the two [Ca ] . m mitochondria were prepared. In intact mitochondria, there was no 2+Prior to solubilization there was no difference in [Ca ] in difference in BECEF fluorescence between treatment groups. m

Following solubilization with Triton X-100, the pH of samples from “locked” mitochondria between treatment groups (Fig. 3). 2+ both DNP- and DMSO-treated animals remained unaltered. These Importantly, background Ca levels from both groups were

2+observations suggest that the difference in [Ca ] between DNP- msimilar to that seen in intact mitochondria from normal, and DMS-treated animals was not due to changes in buffer pH untreated animals (Fig. 3). Following lysis with Triton X-100, following lysis of mitochondria. mitochondria prepared from animals administered DMSO

2+revealed a [Ca ] of 6.99 + 0.136 nmol/mg protein, consistent Acidic environments can cause probes such as Calcium Green 5-m

with what we observed in normal, untreated animals. In N to fluoresce. To determine whether the coincidental release of

A SNRES ARTICLE 52


2+ 13,2protons from the lysed mitochondria affected buffer pH and, that is propels Ca into the mitochondria . In animals hence, florescence intensity, experiments were conducted in the administered DNP, there was an almost 60% reduction in the

2+presence of the pH indicator BECEF. In wells containing concentration of [Ca ] compared to DMSO-treated control m

mitochondria from DMSO- and DNP-treated animals, the pH of animals, consistent with a reduction in mitochondrial the assay was 6.54 + 0.01528 and 6.5567 + 0.1497 respectively. membrane potential. It should be pointed out that DNP may Following solubilization with Triton X-100, there was no change have some non-specific membrane effects, such as those leading

19in the pH of either treatment group, indicating that the change to the release of catecholamines . To control for these effects, 2+in Calcium Green 5-N fluorescence is not due to the liberation of we evaluated the effects of the DNP analogue TNP on [Ca ] . m

19protons from mitochondria (Fig. 4). Like DNP, TNP interacts with the plasma membrane but unlike 14DNP lacks uncoupling activity in intact mitochondria . In

Discussionanimals treated with the molar equivalent of this compound,

2+Mitochondrial calcium homeostasis is critically involved in there was no reduction in [Ca ] indicating the effect of if DNP m2+mechanisms affecting both cell survival and cell death. While on [Ca ] was due to its uncoupling properties and that this m

2+2+limited increases in [Ca ] can activate enzymes in response to m assay is sensitive to changes in [Ca ] in vivo. A limitation to this m

15increased cellular energy demands required for cell survival , assay is that it does not differentiate between free and bound large rises in this same ion can stimulate pathways that result in calcium in the matrix.

1either necrotic or apoptotic cell death . In the central nervous 2+ Although the effect is small, pH can affect the intensity of system, elevations in [Ca ] have been implicated in both acute m

fluorescence following excitation. To be certain that the release neuronal injury, such as stroke and traumatic spinal cord/brain of matrix protons did not change buffer pH, we used BECEF to injury, as well as in neurodegenerative disorders such as measure buffer pH. The lack of change in fluorescence in buffer 3Huntington's disease . The ability to easily and reproducibly pH following lysis of the mitochondria indicates that the 2+quantify in vivo [Ca ] in animal models for these disorders is m changes that we have measured are not a consequence of

thus of great utility. proton accumulation. In light of the observation that the

8Similar to a previously reported method , we have relied on the addition of EGTA to solubilized mitochondrial causes a drop in 2+use of “blockers” to prevent both the influx and efflux of [Ca ] Calcium Green 5-N fluorescence to near baseline levels, we m

during the isolation of the mitochondria, a detergent to rupture conclude that the signal that we are measuring is, indeed, the 2+the mitochondria and liberate calcium and a fluorescent probe result of liberated Ca from the mitochondria.

2+(Calcium Green 5-N) to quantify the liberated Ca . In the Conclusionmethod of Zaiden and Sims, they used ruthenium red to block

2+ the mitochondrial inward transport of Ca via the uniporter and This represents a modified method that allows for the 11extrusion via the sodium-independent antiporter and VDAC . In quantification of in vivo mitochondrial calcium levels. The

addition to ruthenium red, our “locking” buffer included method provides sufficient sensitivity to measure experimentally 16 2+ cyclosporin A to block the mitochondrial PTP . The ability to induced changes in [Ca ] in vivoand should be useful in studies m

block PTP is particularly important for measurements under aimed at determining concentrations of this ion in a number of pathological conditions as it is likely to be activated and may models of neurodegenerative conditions in which altered

2+ 2+allow the flux Ca during mitochondrial preparation. To prevent [Ca ] has been implicated.m 2+ efflux of Ca through the sodium dependent antiporter, Zaiden

+and Sims omitted Na from their “isolation” buffer of. We, too, References+omitted Na however we also added CGP-37157 to prevent 1. Orrenius S., Zhivotovsky B. and Nicotera P. Regulation of cell death: the

17 +activation of the sodium-dependent antiporter by stores of Na calcium-apoptosis link. Nat Rev Mol Cell Biol 2003; 4: 552-565.

located within the mitochondrial matrix. Another difference 2. Gunter T. E., Gunter K. K., Sheu S. S. et al. Mitochondrial calcium between the previous method and the one reported here is the transport: physiological and pathological relevance. Am J Physiol

1994; 267: C313-339.use of chelating agents during tissue preparation. In our 3. Fiskum G., Murphy A. N. and Beal M. F. Mitochondria in method, there was no EDTA/EGTA present in the “locking”

2+ neurodegenera t ion : acu te i s chemia and ch ron i c buffer, thus precluding a possible shift in the equilibrium of Ca neurodegenerative diseases. J Cereb Blood Flow Metab 1999;19: from the mitochondrial matrix into the buffer. Lastly, by using 351-369.

+Calcium Green 5-N rather than the ratiometric dye fura-2K , we 4. Howell S. L. and Tyhurst M. 45Calcium localization in islets of

were able to generate standard curves directly using known Langerhans, a study by electron-microscopic autoradiography. J 2+quantities of Ca . Despite the differences in the two methods, Cell Sci 1976;21: 415-422.

2+the concentrations of [Ca ] in “normal” mitochondria m 5. Borgers M., Thone F. and van Nueten J. M. The subcellular distribution of determined using this modified assay were only slightly higher calcium and the effects of calcium-antagonists as evaluated with a

combined oxalate-pyroantimonate technique. Acta Histochem than that reported previously.Suppl 1981;24: 327-332.

DNP is an uncoupling agent that disconnects the flow of 6. Peng T. I. and Greenamyre J. T. Privileged access to mitochondria of

electrons down the electron chain from ATP synthesis by causing calcium influx through N-methyl-D-aspartate receptors. Mol protons to “leak” back into the matrix form the inner Pharmacol 1998;53: 974-980.

18mitochondrial space . This short circuit results a reduction in the 7. Yuste R. and Denk W. Dendritic spines as basic functional units of mitochondrial membrane potential and, thus, the driving force neuronal integration. Nature 1995; 375: 682-684.

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8. Zaidan E. and Sims N. R. The calcium content of mitochondria from Diego1992.brain subregions following short-term forebrain ischemia and 14. Hanstein W. G. A. H., Y. Trinitrophenol: a membrane-impermeable recirculation in the rat. J Neurochem 1994; 63: 1812-1819. uncoupler of oxidative phosphorylation. Proc Natl Acad Sci U S A

1974; 71: 288-292.9. Sims N. R. Rapid isolation of metabolically active mitochondria from rat brain and subregions using Percoll density gradient 15. Brini M. Ca(2+) signalling in mitochondria: mechanism and role in centrifugation. J Neurochem 1990;55: 698-707. physiology and pathology. Cell Calcium 2003; 34: 399-405.

10. Petronilli V., Cola C. and Bernardi P. Modulation of the mitochondrial 16. Broekemeier K. M., Dempsey M. E. and Pfeiffer D. R. Cyclosporin A is a cyclosporin A-sensitive permeability transition pore. II. The potent inhibitor of the inner membrane permeability transition in minimal requirements for pore induction underscore a key role for liver mitochondria. J Biol Chem 1989;264: 7826-7830.transmembrane electrical potential, matrix pH, and matrix Ca2+. J

17. Cox D. A., Conforti L., Sperelakis N. and Matlib M. A. Selectivity of Biol Chem 1993 ;268: 1011-1016.

inhibition of Na(+)-Ca2+ exchange of heart mitochondria by 11. Tapia R. and Velasco I. Ruthenium red as a tool to study calcium benzothiazepine CGP-37157. J Cardiovasc Pharmacol 1993;21:

channels, neuronal death and the function of neural pathways. 595-599.Neurochem Int1997; 30:137-147.

18. Boveris A. and Chance B. The mitochondrial generation of hydrogen 12. Scanlon J. M., Brocard J. B., Stout A. K. et al. Pharmacological peroxide. General properties and effect of hyperbaric oxygen.

investigation of mitochondrial ca(2+) transport in central Biochem J1973;134:707-716.neurons: studies with CGP-37157, an inhibitor of the 19. Sorimachi M. and Yamagami K. Nitriphenol compound induces Ca-mitochondrial Na(+)-Ca(2+) exchanger. Cell Calcium 2000;28: dependent exocytotic secretion of catecholamines by a direct 317-327. effect on the plasma membranes of the adrenal medullary cells.

13. Nicholls D. G. and Ferguson S. J. Bioenergetics 2. Academic Press, San Brain Res 1982;232:242-246.

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Background have been derived from differentiated pigmented ciliary epithelium and not involve retinal progenitor cells. Hypothesis of Macular degeneration, diabetic retinopathy, glaucoma and this study was to test the efficacy of CE derived stem cells for retinitis pigmentosa are among the major causes of blindness transplantation in patients with retinal degenerative diseases. and visual impairment. Currently available treatment strategies

like laser photocoagulation, photodynamic therapy, anti Study designangiogenesis therapy and drug delivery are ineffective in

1 The study was intended to demonstrate the usefulness of ciliary restoring the normal vision of retinal degeneration patients . epithelium derived neurospheres in restoration of normal vision. Resident stem cells in the eye have a potential for therapeutic use Cicero et al have analyzed morphological features of ciliary to regenerate and repair disease affected retina. The Ciliary epithelium from adult C57BL/6 mice using transmission electron Marginal Zone (CMZ) present in the retina of lower vertebrates microscopy (TEM). Pigmented ciliay epithelium from adult like amphibians, fishes and birds is a source for retinal stem cells

2 C57BL/6 mice was observed to have melanosomes, basal and (RSCs) . The stem cells present in the CMZ have been reported to lateral membrane interdigitations and epithelial cell-cell play an important role in the post natal development and junctions. These features were not found in retinal progenitors regeneration of retina in lower vertebrates. Extensive studies endorsing that adult CE cells and retinal progenitors don't share have been done on the regulation and maintenance of the CMZ

3 any morphological characteristics. Molecular profiling of CE stem cells . The size of CMZ has decreased during the course of revealed that specific genes such as Palm1, Rab27b (which are evolution from amphibians-reptiles-birds and is completely expressed in the fetal and adult pigmented ciliary epithelium) absent in mammals. The lack of CMZ is thought to be responsible were expressed only in the pigmented ciliary epithelium but not for the inability of mammalian retina to regenerate after injury or in the retina. Also, Nrl and recoverin genes are specifically degeneration. Ahmad et al and Tropepe et al reported for the expressed in retina as analysed with real time PCR further first time in 2000 about the existence of a rare population of delineates pigmented CE from retina. CE derived spheres were stem cells in a region of pigmented ciliary epithelium, an

4,5 also examined for the presence of retinal stem and/or progenitor extension of retinal pigmented epithelium (RPE) . These reports cells by RT-PCR (molecular), immunocytochemistry (cellular) and provided hope for the treatment of retinal degenerative diseases TEM (morphological) and then compared with adult CE, retinal through transplantation or mobilization of these cells. The past progenitors and retinal progenitor derived spheres (from P0 decade has witnessed remarkable advances in the proliferation retina). CE derived spheres contained melanosomes, cell to cell and differentiation capacity of ciliary epithelial (CE) stem cells junctions and membrane interdigitations as that of adult and the factors affecting the maintenance of these stem cells. pigmented CE cells which were not found with retinal Proliferation and differentiation potential of CE stem cells has

6 7 8 progenitor cells or retinal neurospheres. Real time PCR analysis been studied in various species like rodents , porcine , rabbits , 9 10 of CE derived spheres revealed that, they share most of their monkey and humans . These cells have also been tested for in

genes with that of adult pigmented CE, but not with the retina. vivo regeneration of retina in various retinal degeneration models. However, it was not clear that in vitro neurospheres To rule out the possibility that sphere are originating from originate from pigmented CE cells itself or involves RSCs or pigmented CE and not from RSCs or progenitor cells present in

orj/orj retinal progenitor cells residing in the CE before Cicero’s et al the CE, authors analyzed CE from Chx10 mutant mice. orj/orj report. In this report, authors have shown that neurospheres Chx10 mutant mice are known to have more number of CE

Cells previously identified as retinal stem cells are pigmented ciliary epithelial cells

a b c d dSamantha A. Cicero , Dianna Johnson , Steve Reyntjens , Sharon Frase , Samuel Connell , Lionel a a e a,b,1M. L. Chow , Suzanne J. Baker , Brian P. Sorrentino , and Michael A. Dyer

a Department of Developmental Neurobiology, TN 38105;b Department of Ophthalmology, University of Tennessee Health Sciences Center, Memphis, TN 38105;c FEI Helios Nanolab, Netherlands;d Cell and Tissue Imaging Shared Resource, ande Department of Hematology, St. Jude Children's Research Hospital, Memphis.

Correspondence : Michael A. Dyer, Deptt. of Developmental Neurobiology & Opthalmology, University of Tennessee Health Science Center, Memphis, TN 38105

Chandrika AbburiNeuroscience Research Lab, Department of Neurology , Post Graduate Institute of Medical Education and Research, Chandigarh

PNAS 2009 ; Vol. 106 : 6685–6690

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cells and are expected to form more number of spheres. TEM Finally, to determine the transdifferentiation capacity of CE analysis of spheres derived from this mutant mouse showed that derived spheres in new borns, cells from CE derived spheres were each cell was pigmented and these spheres were derived from injected into the subretinal space of new born pups and eyes pigmented CE cells rather that RSCs or retinal progenitor cells. To were analyzed after 21 days. They did not find any integration of further confirm these results, authors labeled proliferating cells these cells in the developing retina and injected cells were

3in the sphere with [ H] – thymidine on day 3 for 1hr and then grouped together and formed basal lamina. These results have stained with toluidine blue. When these spheres were analyzed indicated that, CE derived spheres don't have the capacity to by TEM imaging, they showed the morphological characteristics integrate into the retina. But, authors didn't transplant these such as melanosomes, membrane interdigitations and epithelial cells in any retinal degenerative model (don't start a sentence junctions as that of adult pigmented CE. with ). All the results showed that, CE derived spheres do not Although the data indicated that proliferating cells were found share any of its profiles with that of retinal progenitors. As these to be pigmented CE cells, however the authors have not CE derived spheres are pigmented, these cells don't appear to be excluded the possibility of a rare stem cell population. To analyze a good source for transplantation in any retinal degenerative every cell in the CE derived neurosphere, they performed serial disease.electron microscopic analysis and morphometric analysis of

Implicationsentire sphere. Out of 383 cells analyzed, all the cells showed

Characterization of CE stem cells by Cicero et al., yields pigmented CE cell features and none of the cell was non information for future utility of these cells in treating retinal pigmented RSC or neural progenitor cell. Nestin expression was

observed in spheres derived from pigmented CE under degenerative diseases. Though pigmented CE spheres are not proliferation conditions or stem cell medium. These pigmented good source for transplantation authors didn't mention how CE cells may ectopically up-regulate stem cell markers upon and why these cells possess unique proliferation capacity. exposure to stem cell medium despite their differentiatiated Further research is required to achieve transdifferentiatation of state. To distinguish between these two possibilities, authors the CE stem cells into pure retinal stem or progenitor cells which cultured the cells in the presence and absence of stem cell can be used for the treatment the retinal degenerative diseases.medium. During this period, at several time points (0,2,8 and

References 24hrs), cells were tested for nestin expression. Results showed 1. Bylsma G W, Guymer R H. Treatment of age-related macular that, exposure to stem cell medium were sufficient to up-

degeneration. Clin Exp Optom 2005;88(5):322–34.regulate the nestin expression and these nestin expressing 2. Perron M, Harris W A. Retinal stem cells in vertebrates. Bioessays. spheres contained pigment, membrane interdigitations and

2000;22(8):685-8.epithelial juctions similar to that of differentiated CE. It indicates 3. Raymond P A, Barthel L K, Bernardos RL, et al. Molecular characterization that ectopic expression of nestin did not mark stem cell

of retinal stem cells and their niches in adult zebrafish. BMC Dev population in the CE or in the CE derived spheres. Previous Biol. 2006;6:36.studies showed that, there was robust transdifferentiation

4. Ahmad I, Tang L, Pham H. Identification of neural progenitors in the adult between CE, retina and RPE in various species. To test this mammal ian eye . B iochem B iophys Res Commun. hypothesis, authors plated CE derived spheres on laminin coated 2000;270(2):517-21.

coverslips and maintained in serum containing media for 21 5. Tropepe V, Coles B L, Chiasson B J, et al. Retinal stem cells in the adult days. Under these differentiation conditions, each cell retained

mammalian eye. Science. 2000;287(5460):2032-6.pigment and TEM analysis revealed that, the cells retained all

6. Lord-Grignon J, Abdouh M, Bernier G. Identification of genes expressed morphological features of CE cells and none of the retinal in retinal progenitor/stem cell colonies isolated from the ocular

neurons. They also performed RT PCR on cells kept for ciliary body of adult mice. Gene Expr Patterns. 2006;6(8):992-9.differentiation for every 2 days for 21 days. The genes that

7. Gu P, Harwood L J, Zhang X, et al. Isolation of retinal progenitor and stem normally express in the rods (Nrl, Rhodopsin, Gnat1), bipolar

cells from the porcine eye. Mol Vis. 2007;13:1045-57. cells (PKC á) and Muller glia (Carlbp) were not induced

8. Inoue Y, Yanagi Y, Tamaki Y, et al. Clonogenic analysis of ciliary epithelial significantly as that of retina. But, genes like Palm1, Rab27b

derived retinal progenitor cells in rabbits. Exp Eye Res. were increased in these cultures. Along with RT PCR, they have 2005;81(4):437-45.also tested the retinal markers by immunocytochemistry.

9. Jonas J B, Panda-Jonas S, Singh Hayreh S. Retinal progenitor cells in the Majority of cells were positive for cytokeratin, marker for posterior pars plana of rhesus monkeys. Br J Ophthalmol. 2004 pigmented epithelial cells. These results showed that, all the Jun;88(6):836-7.labeled cells retained the pigment and morphology of CE cells. 10. Xu H, Sta Iglesia D D, Kielczewski J L, et al. Characteristics of progenitor Experiments with human CE derived spheres showed same cells derived from adult ciliary body in mouse, rat, and human results as that of murine CE derived spheres. eyes. Invest Ophthalmol Vis Sci. 2007;48(4):1674-82.

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Rene Descartes, a famous mathematician and scientist turned Chandoux that science could only be based on probabilities philosopher, is considered to be the father of modern brought him enormous applause. In 1628, Descartes moved to

Holland and remained there for almost 21 years. During this philosophy, who paved the path to certainty of knowledge period he wrote and published many books – Rules for the through skepticism and said that all knowledge is the product of Direction of Mind (1628/29), Le Monde or The World (written in reasoning based on self-evident assumptions. Dichotomy of 1629), Discourse on Method (1637), Meditations on First mind and body, as proposed by Descartes, was considered to be Philosophy (1641), The Principles of Philosophy (1644), and The the turning point in the modern philosophy, which provided the Passions of the Souls (1649). Descartes' The Principles of basis of the certainty of knowledge and interventionist medical

1 Philosophy was the manuscript of his mature Cartesian system in practice . Evidently, Descartes immortalized his method of general on metaphysics and physics. Descartes' work on the doubting to achieve certainty of knowledge. Passions of the Souls was a combination of psychology,

Descartes was born at La Haye (now called Descartes), France. He physiology and ethics, which contained his theory of two way went to Jesuit College of La Fleche in 1606 where students were causal interaction of mind-body in the pineal gland.trained for career in military engineering, government

Cartesian dualism maintained that soul and body are completely administration and the judiciary. In the journey of his intellectual different kind of substance but they are closely joined and acquisition, Descartes embarked on the field of mathematics,

2intermingled . Descartes gave pain analogy to understand the science, law and philosophy. His mathematical and deductive 3dichotomous interaction of the mind and body . The happiness methodological approach underlined the contours of

and ecstasy felt in the mind on certain news is not the intellectual philosophical reasoning and certainty of the knowledge. His joy, but it sends sensations of joy from brain to heart and excite first substantial work was the Regulae or Rules for the Direction the small nerve there. Similarly, changes in the body can create of Mind written in 1628-29 and published in 1701, which

1sensations in the soul irrespective of its causation . By his reflected his interest in scientific methods. Issac Beechman, a mechanistic view of bodily functions, and interaction with the contemporary mathematician, exerted immense influence on soul, though they being two separate entities, Descartes raised a him and shared some common approach and methods in pertinent question for the medical interventionists of today's dealing with problems of science and mathematics. modern world that – a resolution and/or correct understanding

Descartes was influenced by the deductive methods of of mind-body interaction would provide the basis of the medical mathematics and understood that key to the contemporary interventions. The answer is not simple – though Descartes scientific advances is in the discovery of proper method. The would be considered pioneer and more close to modern thirteen books of Euclid's Elements was a model of deductive biopsychosocial theory of medical intervention.methods for him. However, Galileo Galilei (1564-1642),

Vinod Srivastava, MSW, LCSWDescartes contemporary, was able to synthesize the Clinical Supervisor experimental methods of deductive and inductive reasoning. E.P. Bradley Hospital, Descartes' commitment to scientific methods and certainty of 1011 Veterans Memorial Parkway, knowledge coincided with Renaissance skepticism rooted in East Providence RI -02915 USA

Europe's religious crisis and protestant Reformation. However, E-mail : [email protected] postulated that “I think, therefore I exist”, which proves that there is a thinking being which exists. Even though I Referencedoubt that I don't exist – I cannot doubt a doubter and his having 1. Descartes, R. The Philosophical Writings of Descartes. J. Cottingham, R. sensory perceptions. While “I”, the doubter, is a thinking and Stoothoff and D. Murdoch (trans.), Cambridge University Press,

Cambridge) 1984 : 2. spatially non-extended soul, the body must also exist, which is a 2. Kennington, R. 'Descartes and mastery of nature,' in S.F Spicker (ed.), non-thinking and spatially extended substance responsible for

Organism, Medicine, and Metaphysics, D. Reidel, Dordrecht 1978 : sensory perceptions in that it is non-thinking and spatially 201–223.

extended. Descartes deductively postulated the existence of 3. Duncan G. Mind-Body Dualism and the Biopsychosocial Model of Pain:

mind and body, and its interaction, and provided the basis of What Did Descartes Really Say? Journal of Medicine and 1certainty of knowledge . His confrontations in Paris in 1628 with Philosophy 2000 ; 25 (4) : 488.

RENE DESCARTES (1595-1650)

Mind–Body Dichotomy

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disposed to speak exactly as they believe, but also because very Prefatory Note by the Authormany are not aware of what it is that they really believe; for, as If this Discourse appear too long to be read at once, it may be the act of mind by which a thing is believed is different from that divided into six Parts: and, in the first, will be found various by which we know that we believe it, the one act is often found considerations touching the Sciences; in the second, the without the other. Also, amid many opinions held in equal principal rules of the Method which the Author has discovered,

in the third, certain of the rules of Morals which he has deduced repute, I chose always the most moderate, as much for the from this Method; in the fourth, the reasonings by which he reason that these are always the most convenient for practice, establishes the existence of God and of the Human Soul, which and probably the best (for all excess is generally vicious), as that, are the foundations of his Metaphysic; in the fifth, the order of in the event of my falling into error, I might be at less distance the Physical questions which he has investigated, and, in from the truth than if, having chosen one of the extremes, it particular, the explication of the motion of the heart and of some

should turn out to be the other which I ought to have adopted. other difficulties pertaining to Medicine, as also the difference And I placed in the class of extremes especially all promises by between the soul of man and that of the brutes; and, in the last, which somewhat of our freedom is abridged; not that I what the Author believes to be required in order to greater disapproved of the laws which, to provide against the instability advancement in the investigation of Nature than has yet been of men of feeble resolution, when what is sought to be made, with the reasons that have induced him to write.accomplished is some good, permit engagements by vows and .............Contd. from Previous Issuecontracts binding the parties to persevere in it, or even, for the

PART 3 security of commerce, sanction similar engagements where the purpose sought to be realized is indifferent: but because I did And finally, as it is not enough, before commencing to rebuild not find anything on earth which was wholly superior to change, the house in which we live, that it be pulled down, and materials and because, for myself in particular, I hoped gradually to perfect and builders provided, or that we engage in the work ourselves, my judgments, and not to suffer them to deteriorate, I would according to a plan which we have beforehand carefully drawn have deemed it a grave sin against good sense, if, for the reason out, but as it is likewise necessary that we be furnished with that I approved of something at a particular time, I therefore some other house in which we may live commodiously during bound myself to hold it for good at a subsequent time, when the operations, so that I might not remain irresolute in my perhaps it had ceased to be so, or I had ceased to esteem it such. actions, while my reason compelled me to suspend my My second maxim was to be as firm and resolute in my actions as judgement, and that I might not be prevented from living I was able, and not to adhere less steadfastly to the most thenceforward in the greatest possible felicity, I formed a doubtful opinions, when once adopted, than if they had been provisory code of morals, composed of three or four maxims, highly certain; imitating in this the example of travelers who, with which I am desirous to make you acquainted. when they have lost their way in a forest, ought not to wander

The first was to obey the laws and customs of my country, from side to side, far less remain in one place, but proceed

adhering firmly to the faith in which, by the grace of God, I had constantly towards the same side in as straight a line as possible,

been educated from my childhood and regulating my conduct in without changing their direction for slight reasons, although every other matter according to the most moderate opinions, perhaps it might be chance alone which at first determined the and the farthest removed from extremes, which should happen selection; for in this way, if they do not exactly reach the point to be adopted in practice with general consent of the most they desire, they will come at least in the end to some place that judicious of those among whom I might be living. For as I had will probably be preferable to the middle of a forest. from that time begun to hold my own opinions for nought In the same way, since in action it frequently happens that no because I wished to subject them all to examination, I was delay is permissible, it is very certain that, when it is not in our convinced that I could not do better than follow in the meantime power to determine what is true, we ought to act according to the opinions of the most judicious; and although there are some what is most probable; and even although we should not remark perhaps among the Persians and Chinese as judicious as among a greater probability in one opinion than in another, we ought ourselves, expediency seemed to dictate that I should regulate notwithstanding to choose one or the other, and afterwards my practice conformably to the opinions of those with whom I consider it, in so far as it relates to practice, as no longer dubious, should have to live; and it appeared to me that, in order to

ascertain the real opinions of such, I ought rather to take but manifestly true and certain, since the reason by which our cognizance of what they practised than of what they said, not choice has been determined is itself possessed of these only because, in the corruption of our manners, there are few qualities. This principle was sufficient thenceforward to rid me

Discourse on the method of rightly conducting the reason,and seeking truth in the sciences

Rene Descartes(Reprinted with permission)

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of all those repentings and pangs of remorse that usually disturb the gratification thence arising so occupied my mind that I was the consciences of such feeble and uncertain minds as, destitute wholly indifferent to every other object. Besides, the three of any clear and determinate principle of choice, allow preceding maxims were founded singly on the design of themselves one day to adopt a course of action as the best, continuing the work of self-instruction. For since God has which they abandon the next, as the opposite. My third maxim endowed each of us with some light of reason by which to was to endeavor always to conquer myself rather than fortune, distinguish truth from error, I could not have believed that I and change my desires rather than the order of the world, and in ought for a single moment to rest satisfied with the opinions of general, accustom myself to the persuasion that, except our another, unless I had resolved to exercise my own judgment in own thoughts, there is nothing absolutely in our power; so that examining these whenever I should be duly qualified for the task. when we have done our best in things external to us, all wherein Nor could I have proceeded on such opinions without scruple, we fail of success is to be held, as regards us, absolutely had I supposed that I should thereby forfeit any advantage for impossible: and this single principle seemed to me sufficient to attaining still more accurate, should such exist. And, in fine, I prevent me from desiring for the future anything which I could could not have restrained my desires, nor remained satisfied had not obtain, and thus render me contented; for since our will I not followed a path in which I thought myself certain of naturally seeks those objects alone which the understanding attaining all the knowledge to the acquisition of which I was represents as in some way possible of attainment, it is plain, that competent, as well as the largest amount of what is truly good if we consider all external goods as equally beyond our power, which I could ever hope to secure Inasmuch as we neither seek we shall no more regret the absence of such goods as seem due nor shun any object except in so far as our understanding to our birth, when deprived of them without any fault of ours, represents it as good or bad, all that is necessary to right action is than our not possessing the kingdoms of China or Mexico, and right judgment, and to the best action the most correct thus making, so to speak, a virtue of necessity, we shall no more judgment, that is, to the acquisition of all the virtues with all else desire health in disease, or freedom in imprisonment, than we that is truly valuable and within our reach; and the assurance of now do bodies incorruptible as diamonds, or the wings of birds such an acquisition cannot fail to render us contented. to fly with. But I confess there is need of prolonged discipline Having thus provided myself with these maxims, and having and frequently repeated meditation to accustom the mind to placed them in reserve along with the truths of faith, which have view all objects in this light; and I believe that in this chiefly ever occupied the first place in my belief, I came to the consisted the secret of the power of such philosophers as in conclusion that I might with freedom set about ridding myself of former times were enabled to rise superior to the influence of what remained of my opinions. And, inasmuch as I hoped to be fortune, and, amid suffering and poverty, enjoy a happiness better able successfully to accomplish this work by holding which their gods might have envied. For, occupied incessantly intercourse with mankind, than by remaining longer shut up in with the consideration of the limits prescribed to their power by the retirement where these thoughts had occurred to me, I nature, they became so entirely convinced that nothing was at betook me again to traveling before the winter was well ended. their disposal except their own thoughts, that this conviction And, during the nine subsequent years, I did nothing but roam was of itself sufficient to prevent their entertaining any desire of from one place to another, desirous of being a spectator rather other objects; and over their thoughts they acquired a sway so than an actor in the plays exhibited on the theater of the world; absolute, that they had some ground on this account for and, as I made it my business in each matter to reflect esteeming themselves more rich and more powerful, more free particularly upon what might fairly be doubted and prove a and more happy, than other men who, whatever be the favors source of error, I gradually rooted out from my mind all the errors heaped on them by nature and fortune, if destitute of this which had hitherto crept into it. Not that in this I imitated the philosophy, can never command the realization of all their sceptics who doubt only that they may doubt, and seek nothing desires. beyond uncertainty itself; for, on the contrary, my design was In fine, to conclude this code of morals, I thought of reviewing singly to find ground of assurance, and cast aside the loose the different occupations of men in this life, with the view of earth and sand, that I might reach the rock or the clay. In this, as making choice of the best. And, without wishing to offer any appears to me, I was successful enough; for, since I endeavored remarks on the employments of others, I may state that it was to discover the falsehood or incertitude of the propositions I my conviction that I could not do better than continue in that in examined, not by feeble conjectures, but by clear and certain which I was engaged, viz., in devoting my whole life to the reasonings, I met with nothing so doubtful as not to yield some culture of my reason, and in making the greatest progress I was conclusion of adequate certainty, although this were merely the able in the knowledge of truth, on the principles of the method inference, that the matter in question contained nothing certain. which I had prescribed to myself. This method, from the time I And, just as in pulling down an old house, we usually reserve the had begun to apply it, had been to me the source of satisfaction ruins to contribute towards the erection, so, in destroying such so intense as to lead me to, believe that more perfect or more of my opinions as I judged to be Ill-founded, I made a variety of innocent could not be enjoyed in this life; and as by its means I observations and acquired an amount of experience of which I daily discovered truths that appeared to me of some availed myself in the establishment of more certain. And further, importance, and of which other men were generally ignorant, I continued to exercise myself in the method I had prescribed;

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for, besides taking care in general to conduct all my thoughts acceptable to every one. And yet, that it may be determined according to its rules, I reserved some hours from time to time whether the foundations that I have laid are sufficiently secure, I

find myself in a measure constrained to advert to them. I had which I expressly devoted to the employment of the method in long before remarked that, in relation to practice, it is sometimes the solution of mathematical difficulties, or even in the solution necessary to adopt, as if above doubt, opinions which we discern likewise of some questions belonging to other sciences, but to be highly uncertain, as has been already said; but as I then which, by my having detached them from such principles of desired to give my attention solely to the search after truth, I these sciences as were of inadequate certainty, were rendered thought that a procedure exactly the opposite was called for, almost mathematical: the truth of this will be manifest from the and that I ought to reject as absolutely false all opinions in regard numerous examples contained in this volume. And thus, without to which I could suppose the least ground for doubt, in order to in appearance living otherwise than those who, with no other ascertain whether after that there remained aught in my belief occupation than that of spending their lives agreeably and that was wholly indubitable. Accordingly, seeing that our senses innocently, study to sever pleasure from vice, and who, that they sometimes deceive us, I was willing to suppose that there existed may enjoy their leisure without ennui, have recourse to such nothing really such as they presented to us; and because some pursuits as are honorable, I was nevertheless prosecuting my men err in reasoning, and fall into paralogisms, even on the design, and making greater progress in the knowledge of truth, simplest matters of geometry, I, convinced that I was as open to than I might, perhaps, have made had I been engaged in the error as any other, rejected as false all the reasonings I had perusal of books merely, or in holding converse with men of hitherto taken for demonstrations; and finally, when I letters. considered that the very same thoughts (presentations) which

These nine years passed away, however, before I had come to any we experience when awake may also be experienced when we

determinate judgment respecting the difficulties which form are asleep, while there is at that time not one of them true, I

matter of dispute among the learned, or had commenced to supposed that all the objects (presentations) that had ever seek the principles of any philosophy more certain than the entered into my mind when awake, had in them no more truth vulgar. And the examples of many men of the highest genius, than the illusions of my dreams. But immediately upon this I who had, in former times, engaged in this inquiry, but, as observed that, whilst I thus wished to think that all was false, it appeared to me, without success, led me to imagine it to be a was absolutely necessary that I, who thus thought, should be work of so much difficulty, that I would not perhaps have somewhat; and as I observed that this truth, I think, therefore I ventured on it so soon had I not heard it currently rumored that I am (COGITO ERGO SUM), was so certain and of such evidence had already completed the inquiry. I know not what were the that no ground of doubt, however extravagant, could be alleged grounds of this opinion; and, if my conversation contributed in by the skeptics capable of shaking it, I concluded that I might, any measure to its rise, this must have happened rather from my without scruple, accept it as the first principle of the philosophy having confessed my Ignorance with greater freedom than those of which I was in search. In the next place, I attentively examined are accustomed to do who have studied a little, and expounded what I was and as I observed that I could suppose that I had no perhaps, the reasons that led me to doubt of many of those body, and that there was no world nor any place in which I might things that by others are esteemed certain, than from my having be; but that I could not therefore suppose that I was not; and boasted of any system of philosophy. But, as I am of a that, on the contrary, from the very circumstance that I thought disposition that makes me unwilling to be esteemed different to doubt of the truth of other things, it most clearly and certainly from what I really am, I thought it necessary to endeavor by all followed that I was; while, on the other hand, if I had only ceased means to render myself worthy of the reputation accorded to to think, although all the other objects which I had ever me; and it is now exactly eight years since this desire constrained imagined had been in reality existent, I would have had no me to remove from all those places where interruption from any reason to believe that I existed; I thence concluded that I was a of my acquaintances was possible, and betake myself to this substance whose whole essence or nature consists only in country, in which the long duration of the war has led to the thinking, and which, that it may exist, has need of no place, nor is establishment of such discipline, that the armies maintained dependent on any material thing; so that " I," that is to say, the seem to be of use only in enabling the inhabitants to enjoy more mind by which I am what I am, is wholly distinct from the body, securely the blessings of peace and where, in the midst of a great and is even more easily known than the latter, and is such, that crowd actively engaged in business, and more careful of their although the latter were not, it would still continue to be all that own affairs than curious about those of others, I have been it is. After this I inquired in general into what is essential I to the enabled to live without being deprived of any of the truth and certainty of a proposition; for since I had discovered conveniences to be had in the most populous cities, and yet as one which I knew to be true, I thought that I must likewise be solitary and as retired as in the midst of the most remote deserts. able to discover the ground of this certitude. And as I observed

that in the words I think, therefore I am, there is nothing at all PART 4 which gives me assurance of their truth beyond this, that I see

I am in doubt as to the propriety of making my first meditations very clearly that in order to think it is necessary to exist, I in the place above mentioned matter of discourse; for these are concluded that I might take, as a general rule, the principle, that so metaphysical, and so uncommon, as not, perhaps, to be all the things which we very clearly and distinctly conceive are

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true, only observing, however, that there is some difficulty in composition is an evidence of dependency, and that a state of rightly determining the objects which we distinctly conceive. In dependency is manifestly a state of imperfection, I therefore the next place, from reflecting on the circumstance that I determined that it could not be a perfection in God to be doubted, and that consequently my being was not wholly compounded of these two natures and that consequently he perfect (for I clearly saw that it was a greater perfection to know was not so compounded; but that if there were any bodies in the

world, or even any intelligences, or other natures that were not than to doubt), I was led to inquire whence I had learned to think wholly perfect, their existence depended on his power in such a of something more perfect than myself; and I clearly recognized way that they could not subsist without him for a single that I must hold this notion from some nature which in reality moment. was more perfect.

I was disposed straightway to search for other truths and when I As for the thoughts of many other objects external to me, as of had represented to myself the object of the geometers, which I the sky, the earth, light, heat, and a thousand more, I was less at conceived to be a continuous body or a space indefinitely a loss to know whence these came; for since I remarked in them extended in length, breadth, and height or depth, divisible into nothing which seemed to render them superior to myself, I could divers parts which admit of different figures and sizes, and of believe that, if these were true, they were dependencies on my being moved or transposed in all manner of ways (for all this the own nature, in so far as it possessed a certain perfection, and, if geometers suppose to be in the object they contemplate), I went they were false, that I held them from nothing, that is to say, that over some of their simplest demonstrations. And, in the first they were in me because of a certain imperfection of my nature. place, I observed, that the great certitude which by common But this could not be the case with-the idea of a nature more consent is accorded to these demonstrations, is founded solely perfect than myself; for to receive it from nothing was a thing upon this, that they are clearly conceived in accordance with the manifestly impossible; and, because it is not less repugnant that rules I have already laid down In the next place, I perceived that the more perfect should be an effect of, and dependence on the there was nothing at all in these demonstrations which could less perfect, than that something should proceed from nothing, assure me of the existence of their object: thus, for example, it was equally impossible that I could hold it from myself: supposing a triangle to be given, I distinctly perceived that its accordingly, it but remained that it had been placed in me by a three angles were necessarily equal to two right angles, but I did nature which was in reality more perfect than mine, and which not on that account perceive anything which could assure me even possessed within itself all the perfections of which I could that any triangle existed: while, on the contrary, recurring to the form any idea; that is to say, in a single word, which was God. examination of the idea of a Perfect Being, I found that the And to this I added that, since I knew some perfections which I existence of the Being was comprised in the idea in the same way did not possess, I was not the only being in existence (I will here, that the equality of its three angles to two right angles is with your permission, freely use the terms of the schools); but, comprised in the idea of a triangle, or as in the idea of a sphere, on the contrary, that there was of necessity some other more the equidistance of all points on its surface from the center, or perfect Being upon whom I was dependent, and from whom I even still more clearly; and that consequently it is at least as had received all that I possessed; for if I had existed alone, and certain that God, who is this Perfect Being, is, or exists, as any independently of every other being, so as to have had from demonstration of geometry can be. But the reason which leads myself all the perfection, however little, which I actually many to persuade them selves that there is a difficulty in possessed, I should have been able, for the same reason, to have knowing this truth, and even also in knowing what their mind had from myself the whole remainder of perfection, of the want really is, is that they never raise their thoughts above sensible of which I was conscious, and thus could of myself have become objects, and are so accustomed to consider nothing except by infinite, eternal, immutable, omniscient, all-powerful, and, in way of imagination, which is a mode of thinking limited to fine, have possessed all the perfections which I could recognize material objects, that all that is not imaginable seems to them in God. For in order to know the nature of God (whose existence not intelligible. The truth of this is sufficiently manifest from the has been established by the preceding reasonings), as far as my single circumstance, that the philosophers of the schools accept own nature permitted, I had only to consider in reference to all as a maxim that there is nothing in the understanding which was the properties of which I found in my mind some idea, whether not previously in the senses, in which however it is certain that their possession was a mark of perfection; and I was assured that the ideas of God and of the soul have never been; and it appears no one which indicated any imperfection was in him, and that to me that they who make use of their imagination to none of the rest was awanting. Thus I perceived that doubt, comprehend these ideas do exactly the some thing as if, in order inconstancy, sadness, and such like, could not be found in God, to hear sounds or smell odors, they strove to avail themselves of since I myself would have been happy to be free from them. their eyes; unless indeed that there is this difference, that the Besides, I had ideas of many sensible and corporeal things; for sense of sight does not afford us an inferior assurance to those of although I might suppose that I was dreaming, and that all smell or hearing; in place of which, neither our imagination nor which I saw or imagined was false, I could not, nevertheless, our senses can give us assurance of anything unless our deny that the ideas were in reality in my thoughts. But, because I understanding intervene. had already very clearly recognized in myself that the intelligent

nature is distinct from the corporeal, and as I observed that all Finally, if there be still persons who are not sufficiently

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persuaded of the existence of God and of the soul, by the distinct our ideas might be, we should have no ground on that reasons I have adduced, I am desirous that they should know account for the assurance that they possessed the perfection of

being true. But after the knowledge of God and of the soul has that all the other propositions, of the truth of which they deem rendered us certain of this rule, we can easily understand that themselves perhaps more assured, as that we have a body, and the truth of the thoughts we experience when awake, ought not that there exist stars and an earth, and such like, are less certain; in the slightest degree to be called in question on account of the for, although we have a moral assurance of these things, which is illusions of our dreams. For if it happened that an individual, so strong that there is an appearance of extravagance in even when asleep, had some very distinct idea, as, for example, if doubting of their existence, yet at the same time no one, unless a geometer should discover some new demonstration, the his intellect is impaired, can deny, when the question relates to a circumstance of his being asleep would not militate against its metaphysical certitude, that there is sufficient reason to exclude truth; and as for the most ordinary error of our dreams, which entire assurance, in the observation that when asleep we can in consists in their representing to us various objects in the same the same way imagine ourselves possessed of another body and way as our external senses, this is not prejudicial, since it leads us that we see other stars and another earth, when there is nothing very properly to suspect the truth of the ideas of sense; for we are of the kind. For how do we know that the thoughts which occur not infrequently deceived in the same manner when awake; as in dreaming are false rather than those other which we when persons in the jaundice see all objects yellow, or when the experience when awake, since the former are often not less vivid stars or bodies at a great distance appear to us much smaller and distinct than the latter? And though men of the highest than they are. For, in fine, whether awake or asleep, we ought genius study this question as long as they please, I do not believe never to allow ourselves to be persuaded of the truth of anything that they will be able to give any reason which can be sufficient unless on the evidence of our reason. And it must be noted that I to remove this doubt, unless they presuppose the existence of say of our reason, and not of our imagination or of our senses: God. For, in the first place even the principle which I have already thus, for example, although we very clearly see the sun, we taken as a rule, viz., that all the things which we clearly and ought not therefore to determine that it is only of the size which distinctly conceive are true, is certain only because God is or our sense of sight presents; and we may very distinctly imagine

exists and because he is a Perfect Being, and because all that we the head of a lion joined to the body of a goat, without being

possess is derived from him: whence it follows that our ideas or therefore shut up to the conclusion that a chimaera exists; for it

notions, which to the extent of their clearness and distinctness is not a dictate of reason that what we thus see or imagine is in

are real, and proceed from God, must to that extent be true. reality existent; but it plainly tells us that all our ideas or notions

Accordingly, whereas we not infrequently have ideas or notions contain in them some truth; for otherwise it could not be that

in which some falsity is contained, this can only be the case with God, who is wholly perfect and veracious, should have placed

such as are to some extent confused and obscure, and in this them in us. And because our reasonings are never so clear or so

proceed from nothing (participate of negation), that is, exist in complete during sleep as when we are awake, although us thus confused because we are not wholly perfect. And it is sometimes the acts of our imagination are then as lively and evident that it is not less repugnant that falsity or imperfection, distinct, if not more so than in our waking moments, reason in so far as it is imperfection, should proceed from God, than further dictates that, since all our thoughts cannot be true that truth or perfection should proceed from nothing. because of our partial imperfection, those possessing truth must But if we did not know that all which we possess of real and true infallibly be found in the experience of our waking moments proceeds from a Perfect and Infinite Being, however clear and rather than in that of our dreams.

.....to be continued in next issue of ANS

Neuroscience Research Lab, Department of Neurology, Post Graduate Institute of Medical Education and Research, Chandigarh, INDIA

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A SNCOMP REVIEW

Neel Kamal Sharma, Sudesh Prabhakar, Akshay Anand

ABSTRACT

Age related macular degeneration (AMD) is the most common cause of visual impairment and is characterized by drusen formation, geographic atrophy and gradual loss of vision. It is a frightening disease that destroys the macula, the central part of retina, severely regimenting a person's normal sight. The utility of mouse models with features of AMD along with its recently reported association with complement factor H-gene, and TLR 4 genes strongly suggest the importance of inflammatory mediators and complement in the pathogenesis of this disease. Current treatment modalities include photodynamic therapy and photocoagulation, however, their efficacy is limited. The animal models that faithfully replicate the features of human AMD are useful platforms in validating new therapies. This not only provides new insights for preventative and restorative approaches in AMD in future but also advances our understanding of cardiovascular and neurodegenerative disorders.

KEY WORDS : Macular degeneration, Risk Factors, Genes responsible for AMD, Genetic predisposition, Animal models, Therapeutic approaches.

Corresponding Author : Akshay Anand, E-mail : [email protected], Tel. : +91-172-2756090

Introduction preceded by drusen formation. Drusen are characterized by progressive apoptosis of amorphous deposits that form between cells in the epithelial layer, geographic Any impairment in vision severely limits the retinal pigment epithelium (RPE) and atrophy in the overlying photoreceptor the ability to maintain a good quality of Bruch's membrane and comprise immune cells and in the underlying cells in the life. Age-related macular degeneration

4deposits, lipofuscin and complement. It is choroidal capillary layer . Wet AMD looks (AMD) is a leading cause of irreversible the spatiotemporal confluence of these quite different in this form with blindness in people over 60 years of age in drusen that partly contributes to vision proliferating blood vessels hemorrhage many industrialized countries. It is the loss. that exude serous fluid under the retina. principal cause of irreversible, registered

1 These new vessels usually form a It is also interesting to note that age-legal blindness on three continents . It is neovascular membrane that originates related disorder shares many mechanisms characterized by the progressive degene-from the choroids and penetrates Bruch's common to atherosclerotic plaque ration of the retina, RPE and underlying

5membrane into the subretinal space . development and drusen formation in choroid (the highly vascular tissue 2 Current therapeutic efforts and clinical glomerular nephritis . Drusen deposition beneath the RPE) which results in severe

trials are directed towards hailting the is a significant risk factor for progression vision loss. The earliest clinically visible growth of the neovascular membrane in to CNV, the exudative hallmark of late abnormality in AMD is the accumulation wet AMD, using angiogenesis inhibitors AMD, and vision loss. Existing animal of drusen (lipoproteinaceous deposits) in such as anti-VEGF or thermal lasers. models attempting to simulate AMD the extracellular matrix between the RPE

through high-fat diets and phototoxicity, Despite considerable research, little is and the choroid. The affected portion of senescence acceleration or candidate known about the events that trigger AMD. the eye is the centre of the retina, called

1,3 gene manipulation do not fully replicate Drusen deposition precedes both the dry the macula, which contains the highest the clinical, histologic and angiographic and wet forms and is an independent risk concentration of cone photoreceptors features of the human condition. factor for the development of a and is responsible for central vision.

6neovascular membrane . Drusen are Although the majority of AMD cases are Recent evidence suggests that comple-considered by some to be the residue of the less-severe dry form, ~10– 20% of ment activation and immune complex undigested material from dysfunctional these patients develop wet, or exudative, deposition occurs in eyes of humans with phagocytic cells in the epithelial layer. AMD. The dry form is characterized by AMD. In other immune complex They seem to consist of general biological geographic atrophy, the formation of deposition disorders, it has been effluvia cholesterol-rich lipids, various pigmented yellow deposits (called drusen) postulated that these proteins serve as an proteins and the like. In particular, and the gradual loss of central vision, in f lammatory focus by inc i t ing oxidized lipoproteins have been identified whereas the wet form of AMD manifests macrophage recruitment through Fc and

7in these structures supporting the notion with rapid development of choroidal complement receptor binding, triggering that they arise through oxidative damage. neovascularization, subretinal membrane humoral activation and phagocytosis. Dry They also contain potential macrophage formation and photoreceptor atrophy, AMD is much more common than wet

leading to blindness. CNV is characterized AMD, but the wet form occurs in about chemoattractants, such as complement 8by bleeding or fluid leakage and is usually 10% of these cases. Dry AMD is components and immunoglobulins.

Age related macular degeneration – advances and trends

A SNCOMP REVIEW63


AMD emerges during a preclinical, asymptomatic phase in which waste material (drusen) accumulates in the space between the basement membrane (Bruch's membrane) and the epithelial layer. Bruch's membrane separates the retina from the choroids, the vascular layer of the eye wall that lies between the retina and the sclera. The epithelial layer, dubbed the retinal pigmented epithelial layer is crucial for photoreceptor health. Cells in this layer recycle visual pigment (rhodopsin), phagocytose photoreceptor tips daily as part of rod and cone regeneration and transport fluid across the membrane to the choroids to help prevent detachment of the neural retina. The metabolic demands on the retinal ep i the l ium are cons iderab le as phagocytosis of photoreceptor tips occurs each morning on awakening for 30 to 60 minutes. Central vision vanishes when

epithelial layer, constitutively produce choroidal vasculature, similar in function cells in this epithelial layer cease to to astroglia in the central nervous system MCP-1 and can secrete high levels of the function properly, causing photoreceptor

19(CNS) . The cells lie in a monolayer molecule after exposure to other degeneration. coupled by a variety of cell–cell junctions cytokines and chemokines such as IL.

The clinical studies show a correlation between RPE and choroidal endothelial MCP-1 might help coordinate the turnover between drusen format ion and 20cells . The apical surface of the RPE of resident macrophages and surveillant, autofluorescence (as seen through a ensheathes the photoreceptors in the immature and surveillant, immature 9scanning laser ophthalmoscope ), an adult retina and supports the renewal of dendritic cells, both of which form indirect measure of the accumulation of their outer segments. Signalling between extensive cellular networks in the choroids pellet-like deposits in the retinal epithelial the retina and the RPE is complex. RPE that intimately contact with the epithelial cell layer. These deposits, called lipofuscin

13 apical membranes contain ion pumps, cell layer . granules, are partially degraded cell lectin affinity sites, transporters for water membranes in lysosomes, and accumulate and neuroactive peptides and receptors RPE – the strategic player in cells that endocytose oxidized for catecholamines and sugars. The

The neuroectodermally-derived RPE is a lipoproteins but have no mechanism to macrophage capacity of RPE cells is major component of the blood–retina adequately dispose of them. This believed to support the maintenance of barrier lying adjacent to Bruch's deficiency occurs in professional photoreceptors and the clearance of the

10 membrane, of which the decreased lipoprotein scavengers , such as the aging complement 5 (C5) and IgG deposits that collagen solubility with age is proposed to 21macrophages in atherosclerotic lesions comprise drusen . It is unclear whether 14contribute to debris accumulation . AMD (foam cells) and in nonprofessional the endogenous macrophages that is partially characterized by the disruption phagocytes, such as neurons in the central accumulate in the choroids of patients of interactions of the epithelium with the nervous systems and cells in the retinal with AMD serve protective or destructive neural retina and the underlying choroidal epithel ia l layer. Defects in this functions. Although they might aid

22vasculature. Retinal function is signifi-housekeeping function in retinal epithelial drusen clearance , they could also 23cantly dependent on the health of RPE ce l l s and in res ident choroidal promote CNV , similar to their proposed

24,25cells, which produce useful cytokines and macrophages may contribute to drusen role in a laser-injury mouse model . 11 maintain the homeostasis of essential accumulation with age . In addition to its These findings suggest that macrophages

photopigments. In the normal retina, the well-known pro-inflammatory talents, can serve both pro- and anti-inflammatory RPE performs the functions of barrier, MCP-1 may also have an immunomo- capacities and that the dysregulation of macrophage and neuroprotective cell dulatory role. For instance, it seems to clearance functions might facilitate layer. The RPE cell layer forms a polarized shift the balance of the T-cell immune d r u s e n f o r m a t i o n a n d d i s e a s e and selectively permeable barrier and response towards T-helper type 2 by progression. These drusen might lead to

15 16,17 controls shape , ion flux and cell disturbances in spatiotemporal sensitivity, promoting production of the cytokine 18 signalling . The basal surface of the RPE colour contrast and central vision. Most of interleukin-4 (IL-4), and by suppressing

sits on Bruch's membrane, which is a these cells are post-mitotic and last for the the T-helper type 1 associated cytokine IL-12 complex extracellular matrix thought to duration of a human life. In this state, the 12 cells of the blood retinal barrier,

be the product of both RPE cells and the cells make and secrete retino-protective including those of the retinal pigmented

Retinal architecture

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A SNCOMP REVIEW

41and anti-angiogenic factors, such as in Caucasian populations . These genetic inflammatory role in the disease. The pigment-epithelium-derived factor studies were recently extended by the a n a l y s i s o f t y r o s i n e - h i s t i d i n e

26,27(PEDF) and fibroblast growth factor observation that polymorphisms in other polymorphisms, primarily involved in CFH, 28(FGF-2) . They also express Fas ligand should also be studied in other complement genes, notably those coding

(FasL), and together these factors are populations. Not withstanding this for factor B-complement component C2 reported to control the growth and report, the genetic basis of AMD as a (BF-C2) and complement C3 (C3), are also

42,43development of new subretinal vessels, single-gene disorder is uncertain. The associated with AMD . There have been 29which can damage vision ; however, their genes encoding chemokine ligand-2 studies where LOC387715 and HTRA1

role in AMD has not been conclusively (Ccl2) and its cognate receptor, polymorphisms have been associated with 30established . chemokine receptor-2 (Ccr2), have also a higher risk of exudative AMD in northern

evoked considerable interest after a new Chinese although there are also reports Melanin is another principal pigment in animal model for AMD was developed. where no association of CFH Y402H with RPE that accumulates only in the

44 Gene hunters are screening the Ccl2 and exudative AMD have been suggested . nondividing cells of the RPE layer. Mature Ccr2 genes in patients with AMD through AMD susceptibility, like any disease, is melanin is known to act as an efficient various multi-centre studies. It is unclear if linked to both genetic and environmental antioxidant which scavenges free radicals

32,33 the mirage of Ccl2 or Ccr2 polymorphisms factors, although its precise etiology and protects the eye from stray light . 60would provide any causal link to AMD . remains elusive. Reported risk factors White mice are, therefore, expected to be

However, current studies among include ocular pigmentation, dietary more susceptible to neovascularization 34 Caucasian populations and US population factors, positive family history for AMD, than black mice . When RPE cells are

did not provide any evidence of an smoking, and several gene mutations such injured by mechanical or thermal damage, association between TLR4, CCR2, and as ATP-binding cassette transporter the cells respond with regeneration, CCL2 and AMD, which implies that the protein 4 (ABCA4), apolipoprotein E fibrovascular proliferation and the

45-5035,36 common genetic variation in these genes (APOE), and fibulin-5 (FBLN5) . mobilization of melanin pigment . does not play a significant role in the Moreover, genome-wide linkage studies Lipofuscin, which is chiefly composed of

61 etiology of AMD although suggested by have successfully identified several major the fluorophore A2E (N-retinylidene-several other studies. CFH polymorphism chromosomal regions including 1q31 and Nretinylethanolamine), accumulates as a

51-55 Tyr402His among Indian patients appears 10q26 . Furthermore, studies of Hong hydrophobic component of RPE in AMD-62indicative of AMD pathogenesis .Kong Chinese and Caucasian population affected eyes and is believed to originate

have identified SNP rs11200638:G>A in from the incomplete lysosomal digestion Kaur et al recently reported the the promoter of high-temperature of shed photoreceptor outer segments, association of LOC387715 and HTRA1

which increases proportionately with requirement A-1 (HTRA1 ) gene, SNPs, along with their risk estimates 37,38age . Thus, the preservation of RPE approximately 6.1 kb downstream of among Indian patients with AMD which

functions and compensation for age- r s 1 0 4 9 0 9 2 4 i n LO C 3 8 7 7 1 5 o n emphasize their significant involvement in related changes are central to the chromosome 10q26, to be associated AMD susceptibility, which may be useful

56,57 63prevention of AMD. Although newer with increased risk of AMD . There are for predictive testing . The role of medical approaches, such as PDT, the studies to suggest that theTyr402His complement in laser-induced CNV has

39 64injection of angiostatic steroids or laser coding allele is associated with wet AMD been suggested recently . This might 58photocoagulation, attempt to inhibit the in the Israeli population . With several loci explain the Janus-like context-dependent

growth of abnormal blood vessels from being implicated in the pathogenesis of role of macrophages: the clearance of the choriocapillaries into the outer retina, AMD, another group has identified that immune deposits at one time and CNV at there are greater challenges for the one of the genetic variants (251A/T), other. The role of multigene effects in development of novel gene and cell which is associated with a gene that AMD is conspicuous from many published

65therapies to treat the causes of this retinal boosts the production of interleukin 8 reports. Zareparsi S et al. have studied disease. (known as IL-8), was significantly more the susceptibility of patients with AMD

common among the patients with AMD. who carry the variant of Toll-like receptor 4 Genetic predisposition This held true even after taking account of TLR-D299G and found it to have an Studies on the molecular composition of age, sex, weight, and smoking, which is a additive effect to AMD, in contrast to its

59drusen have implicated inflammation, known risk factor for AMD. Klein et al susceptibility pattern in atherosclerosis. In and particularly local activation of the have reported a genome-wide screen of contrast Edwards et al found that TLR3, alternative pathway (AP) of the 96 ca se s and 50 cont ro l s fo r TLR4, and TLR7 were unlikely to have a

66complement cascade in the retina, in the polymorphisms in the complement factor major impact on overall AMD risk and 40pathogenesis of AMD . Furthermore, H gene (CFH) that are associated with hence the deeper investigation into the

strong evidence for a role of complement AMD. In individuals that are homozygous role of different genes in different in this disease derives from an for the CFH risk allele, the likelihood of populations can not be ruled out. A report independent line of research which AMD is increased by a factor of 7.4. This of the predominance of an Alu-sequence showed that variants in the complement report tends to prove the 'common-gene– insertion in the angiotensin-converting factor H (CFH) gene are significantly common-disease ' hypothesis and enzyme (ACE) showed it to be protective associated with an increased risk for AMD endorses the strong association of an in unaffected individuals and can be

A SNCOMP REVIEW65


exploited as a plausible target for the dry blood flow around the eye or affecting the Animal Models67 68 72form of AMD . Fiotti and co-workers pigments (coloration) in the retina . AMD Progress in the field of AMD has been slow

doubles the risk of dying from a heart have reported a positive correlation because of a lack of animal models. The attack or stroke, reveals research between the existence of longer satellites recent development of animal models for published in the British Journal of (CA) in the promoter region of the gene AMD has allowed researchers to 73Ophthalmology . In the normal retina, encoding matrix metalloproteinase-9 investigate more translational and RPE performs the funct ions of (MMP-9) and the exudative form of AMD, innovative therapies and pathogenetic macrophages and a neuroprotective cell 76,77prompting further analysis of complex studies of AMD . Many animal models layer. During aging and with the interactions between different genes have been used to simulate AMD through progression of the disease, the degree of 78,79implicated in AMD. In addition, raised high-fat diets and phototoxicity , each of these functions diminishes. As a 80levels of MMP-9 in plasma revealed the senescence acceleration or candidate-

69 result of a complex series of molecular 81-83association with AMD patients . Elevated gene manipulation , resulting in partial events, including environmental, serum/plasma levels of CRP have also been clinical, histological and angiographic 70 biochemical and genetic factors, AMD associated with neovascular AMD features of the human condition. In mice develops. It is chiefly accompanied by

fed with high-fat diets, Dithmar and Several studies have investigated the aberrant regulation of the CFH gene coworkers reported age-dependent superoxidedismutase gene polymorphism (complement factor H) in cells and higher electron-lucent debris and in AMD and found no significant complement 5 (C5), immunoglobulin G cholesterol levels compared with those contribution towards its progression (IgG), tissue inhibitor of metallo-fed normal diets. There are some unlike other progressive disorders. The proteinase-3 (TIMP-3) and lipofuscin similarities with basal linear deposits, direct role of VEGF, fibroblast growth deposit formation beneath the RPE, which although the debris does not form a factor-2 (FGF 2) and angiopoietins in CNV facilitates the induction of neovasculari-discrete layer external to the basement has been extensively studied and zation and the consequent blurring of membrane of the RPE, which occurs in established even though there are isolated vision. With the recent postulation of an basal linear deposits. This animal model of reports of FGF2 gene not inhibiting CNV in association of the CFH gene with AMD and Blam-D-like material in Bruch's membrane the laser-injury mouse model. An intricate characterization of chemokine-deficient is similar to the deposits that occur in AMD balance of pro- and anti-angiogenic knockout mice with features resembling

3 but many other clinical features, such as moieties, therefore, is believed to be AMD , considerable excitement has been d r u s e n , i m m u n e d e p o s i t s a n d responsible for determining the status of generated in this field. Recent study photoreceptor atrophy, are lacking. Some CNV. For example, angiotensinogen (Ang) among AMD patients and controls has eyes in this older group also develop I and Ang II impart pro-angiogenic and Tie revealed elevated plasma concentrations endothelial invasion into Bruch's of activation products C3d, Ba, C3a, C5a, 2 (a receptor tyrosine kinase) and PEDF membrane but none of these has classic SC5b-9, substrate proteins C3, C4, exert anti-angiogenic effects, inhibiting

79linear deposits. Cousins and co-workers regulators factor H thus providing CNV. As expected, aged RPE has reduced evidence for an association of systemic have demonstrated that age increases the expression of PEDF; however, it is not clear activation of the alternative complement capacity of dietary fat, especially in the if this is due to its decreased synthesis or pathway with genetic variants of CFH that presence of environmental light, to induce increased proteolytic activity in the were previously l inked to AMD sub-RPE deposits, but these studies vitreous.

74susceptibility . Surprisingly, smoking provide little information about the Risk Factors intensity had reported to have a negative understanding of pathophysiology of

80impact on general-, near-, and far visual AMD. Majji et al. have described Genetic factors, age, cigarette smoking, functional status independent of AMD- senescence-accelerated mice (SAMP8) as nutrition, and exposure to light have been status. Quitting smoking seemed to have a models that show choriocapillary atrophy, identified as AMD risk factors. According time-dependent protective effect on near abnormal deposits in Bruch's membrane to the prevalence estimates from World

75and far vision . and intra-Bruch's membrane CNV, Health Organization AMD is the most although this model doesn't capture most common cause of blindness in developed Research to understand the dynamics of of the features of AMD. Nevertheless, countries and it is estimated that 8 million RPE biology during aging has stimulated given the rare occurrence of CNV in mice, people are blind worldwide or severely interest in preventive and prophylactic the study of angiogenic processes will visually impaired as a result of disease. The therapies for earlier intervention during provide insights into the secondary effects prevalence of AMD, already high in the degenerative processes. At present, of CNV on photoreceptor disruption, developed countries, is likely to increase laser photocoagulation and/or photo-which ultimately leads to deficits in visual dramatically according to some estimates dynamic therapy (PDT) with steroids is

71 perception. The dystrophic RCS (Royal by 50% with aging of the population . known to slow the progression of the College of Surgeons) rat is another model Smoking has been identified as one of the disease temporarily and is the only that undergoes progressive photorecep-few modifiable risk factors for AMD, a treatment available, but with better tor degeneration due to a primary defect leading cause of vision loss in older understanding of these molecular in RPE cells. Although it does not present Americans. Smoking may contribute to mechanisms, advanced therapeutic

AMD through several pathways, including approaches can bring hope to patients with spontaneous CNV, it is useful for reduction of antioxidant levels, decreasing with AMD. assessing the responsiveness of RPE

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t r a n s p l a n t a t i o n a n d s t e m - c e l l function by electrooculogram. Although mice, lending support to the role of 84,85implantation studies . there are several reports of immune- complement participation in CNV and

mediated processes leading to drusen AMD. Combined with the advancement of The use of VEGF-overexpressing mice 91-93 1formation , Ambati and Anand our understanding of a causal link under the control of the RPE65 promoter

described the first animal model for the between the gene encoding complement is a potential tool for analysing 86 development of drusen and lipofuscin factor H and the pathology of AMD , it has intrachoroidal neovascularization . This

deposits in the RPE or spontaneously become possible to organize the model might be useful to screen for occurring CNV resembling that seen in inflammatory paradigm in AMD into a inhibitors of choroidal vessel growth. The patients with AMD. These features more cogent concept. With such currently used laser-injury model for develop as the Ccl2-and Ccr2-knockout developments, there is likely to be a major studying CNV, postulated by Ryan et al. is mice age. Ccl2 is an 8–10-kDa chemotactic shift in our approach to the treatment of another model that recapitulates some of cytokine that recruits monocytes to this disease. Recent study investigated the cardinal features of CNV to which inflammatory sites. It acts through a Ccl2-/-/Cx3cr1-/- [double-knockout (DKO)] many drug development studies have

87 transmembrane-domain G-protein- mice developed AMD-like retinal lesions been adapted . There is a small fraction of 94 such as abnormal retinal pigment coupled receptor called Ccr2 . RPE cells belly spot and tail heterozygote knockout

+/- epithelium cells, drusen, photoreceptor secrete Ccl-2 in a polarized fashion (Bst ) mice that spontaneously develop 95 atrophy and choroidal neovascularization, through their basal surface . Through in CNV. However, the majority of these

which progressed with age and reversed vitro and in situ studies, Ambati and co-heterozygous mice exhibit other severe 88 with h igh omega-3 long-cha in workers demonstrated that choroidal ocular disturbances and therefore their

polyunsaturated fatty acid diet. A broad macrophages migrate by Ccl-2 gradient, application for AMD might be limited. +/- spectrum of AMD pathologies with early adhere to and degrade the immune Nearly 60% of Bst mice have abnormal

onset and high penetrance in these mice deposits. Ccl-2- and Ccr-2-deficient mice pupillary light responses owing to implicate certain chemokines, A2E and have an impaired recruitment of unilateral or bilateral atrophy of the optic endoplasmic reticulum proteins in AMD macrophages. It has been suggested that, nerves. The ocular phenotype of patients

98pathogenesis .as a result of this dysfunction, there is an with early or late AMD bears no accumulation of complement fragments resemblance to the panoply of structural

Current therapeutic approaches – +/- that might damage RPE and induce VEGF abnormalities in Bst mice, which changing concepts production by these cells, resulting in the probably result from global embryonic

development of CNV. Although the Current therapies either target RPE or CNV. dysfunction rather than the specific possibility of macrophage recruitment The wet, or exudative, form of macular perturbation of cellular pathways. It is that is independent of the Ccl-2–Ccr2 axis degeneration affects 15% of those known that TIMP-3 has an important role has not been excluded, such as through diagnosed with AMD, but accounts for in CNV. Sorsby fundus dystrophy (SFD) is a RPE trophic or anti-angiogenic factors, ~80% of the visual complications leading rare late-onset macular dystrophy caused these eyes exhibit the morphological, to blindness. Conventional laser-by mutations in the TIMP3 gene. Weber

89 ultrastructural and functional features photocoagulation surgery to remove CNV and co-workers generated knock-in mice that are characteristic of human AMD. The is indicated in only 10–20% of patients displaying early features of age-related occurrence of CNV is subdued (in ~25% of with wet AMD. After laser surgery, changes in Bruch's membrane and the Ccl2- and Ccr2 knockout mice) but the patients report diminished visual acuity. RPE. These are good for studying the role detection of C5, IgG, TIMP-3, SAP, Likewise, PDT, which is used to halt the of metalloproteinases in the pathogenesis advanced-glycation end products (AGEs), growth of blood vessels into the central of neovascularization. These mice, A2E (a component of lipofuscin) and VEGF retina, is effective in a small proportion of however, take a long time to display by immunostaining and western blotting patients with wet AMD. Tiwari et al have abnormalities in the inner aspect of is in reasonable consonance with the reported that the visual outcomes of Bruch's membrane and the organization features of AMD pathology. These photodynamic therapy (PDT) with of the adjacent basal microvilli of the RPE. pathologies are absent in age-matched v e r t e p o r f i n a n d t r a n s p u p i l l a r y Another model for is the best vitelliform wild-type mice and several other knockout thermotherapy (TTT) for classic subfoveal macular dystrophy (VMD) model in rats strains of mice that were analysed, such as choroidal neovascularization (CNVM) and has been recently proposed for macrophage inflammatory protein (MIP secondary to age-related macular studying AMD because it shares common

- /- - /-90 1á )-, B5 integrin (B5 )- and Chemokine degeneration (ARMD) in one clinical trial. features with AMD . Bestrophin, which is - /-receptor 5 (Ccr5 )-knockout mice. Even T h e y o b s e r v e d t h e s h o r t - t e r m a 68-kDa basolateral plasma-membrane

- /-in a study of apolipoprotein E (apoE preservation of vision in patients of classic protein that is expressed in RPE cells, is a 2+ mice), BlinD and BlanD were reported to CNVM due to ARMD, PDT seems to be regulatory component of Ca channels

be elevated but no drusen or CNV were better than TTT if the pre-laser best and is encoded by the VMD2 gene, which 96 97observed . Bora and co workers have corrected visual acuity is > 20/63 but both is mutated in Best macular dystrophy. This

recently recapitulated some elements of are equally effective if pre-laser best model is characterized by deposits of 99the Ccl-2 and Ccr-2 model by reporting a corrected visual acuity is < 20/63 . lipofuscin in Bruch's membrane with

accompanying photoreceptor atrophy reduction in laser-induced CNV in cobra- Clinicians observe that abnormal blood-- /-and can be used for studying the RPE venom-depleted complement and C3 vessel growth reoccurs after either laser or

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100,101PDT . Clearly, additional therapeutic demonstrate that repeated protein tested for its anti-angiogenic effects, but options are needed to address this production in the eye can be stimulated was not promising because of strong side

114,115problem and to restore the lost function. from the cytomegalovirus (CMV) effects . Genistain has also been Considering the known effects of promoter without repeat intraocular studied to inhibit choriocapillaries

107 116oxidative stress on RPE cell health and injections using a small molecule , all- regeneration . Perhaps there is a need to function, an obvious therapeutic trans retinoic acid (ATRA), which is by shift the focus from a destructive (CNV approach for the maintenance of RPE cell systemic delivery can stimulate protein resolution) to a restorative paradigm. The function is nutritional supplementation production multiple times in the eye and new treatment strategy of reconstruction with antioxidants. The value of resulted in stimulation of gene expression of the altered subretinal structure by antioxidant supplements in slowing the to relevant levels that block abnormal maculoplasty (macular reconstruction) progression of age-related degenerative blood vessel growth in an experimental might be helpful. This serves to replenish a changes has recently been supported by animal model for AMD. These data normal photoreceptor–RPE interface. long-term studies such as the Age-Related suppor t the p r inc ip l e s o f th i s Some groups have been attempting the

102,103 + Eye Disease Study (AREDS) . Clinical technological discovery to therapeutic use of bone-marrow-derived CD34 cells studies of dietary supplements that applications for chronic ocular diseases. for regeneration of the retina in contain carotenoids, anti-oxidant Strategies for treating AMD with gene experimental animal models. It remains to vitamins A, C and E and minerals such as transfer are being developed to address be seen how beneficial the effects are zinc show a 25% decrease in the rate of the dilemmas of systemic toxicity, immune going to be. So far, no clear consensus of progression to aggressive AMD among reaction and nonspecific cellular appropriate genetic targets for either dry high-risk patients. Similar in scope to expression. Disease-causing mutations in or wet AMD has been elucidated. The vitamin supplementation and sub- genes (e.g. CFH, ATP-binding cassette restoration of visual function is a central threshold laser, these therapies have the receptor (ABCR), TIMP3 and Ccl2 and component for the development of a potential for broad use because they can Ccr2) that regulate interactions between clinically acceptable cell or gene therapy be effective as preventive or prophylactic the RPE and retina in two of the early- for AMD. The potential for a rescue-of-treatments. onset forms of macular degeneration function effect of subretinal viral delivery

might be potential targets for gene of the Ccl2 and Ccr2 or CFH genes coupled Considerable progress has been made 108,109therapy . An improved understanding with stem-cell therapy might be the with experimental cellular and gene

of the role of vascular endothelial growth rationale replete with the greater hope. It therapies for macular degeneration. The factor (VEGF) in the genesis of choroidal is uncertain whether this would revive the ultimate goal of experimental RPE cell neovascular membranes has led to the appropriate number of specialized transplantation is to supplement the RPE creation and use of intravitreous anti-cell layer with healthy native or genetically macrophages at the site of drusen VEGF antibodies (bevacizumab and engineered cells to prevent further deposits, to participate in drusen ranub i zumab) and an ap tamer atrophy of the RPE and retina. degradation without relieving the (pegaptanib) in the treatment of these Transplantation has been performed in inhibition of the CFH complement lesions. These new intravitreous injections animal models and in limited human pathway in RPE. Although, recently the for AMD have supplanted previous clinical trials; however, the desired implantation of neural retinal progenitor treatments in both efficacy and safety and restoration of normal function has not cell layers (sheets) with its retinal pigment

104-106 are now the standard of care for been conclusively demonstrated . epithelium (RPE) in dry AMD showed neovascular AMD. Many clinical trials have Recently, Radtke et al. demonstrated the improved visual acuity scores in 70% adopted an anti-VEGF approach as a efficacy and safety of the implantation of patients. This outcome provides clinical standard target for AMD. These aim to neural retinal progenitor cell layers in evidence of the safety and beneficial

31curtail the abnormal blood-vessel AMD patients , out of which 70% patients effect of retinal implants .Treatment formation that obliterates vision. A exhibited improved visual acuity. Thus this options for wet AMD are still at a relatively synthetic VEGF-aptamer RNA binds to outcome provides clinical evidence of the early stage of evaluation and at present no extracellular VEGF and tends to inhibit the safety and beneficial effect of retinal treatment for dry AMD is routinely CNV. Even soluble fms-like tyrosine kinase implants and corroborates results in available.(sflt-1) and angiostatin delivery has been animal models of retinal degeneration.

Paradoxesattempted in an effort to contain the Gene therapy to modify the expression of 110CNV . The clinically relevant inhibition of In spite of rapid strides in this field, the proteins in the RPE cell layer has the

the protease system (uPA–uPAR) might spectrum of genetic targets involved in p o t e n t i a l t o p r e v e n t m a c u l a r prove to be a potentially novel anti- the pathogenesis of AMD has become degeneration at the molecular level. angiogenic therapy for CNV if a recent more complex . Determin ing an Delivery of therapeutic proteins, such as report in a laser CNV model is to be appropriate target in this disease is the antiangiogenic proteins, to the eye is a

1 1 1believed . Intercellular adhesion biggest obstacle. Because most of the demonstrated method for the control of molecule (ICAM-1) blockade has also been information is derived from animal age-related macular degeneration (AMD).

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489–499. neovascularization by a peptide 121. Anand A, Skufca D W, Ambati B K et al. inhibitor of the urokinase Vascular endothelial growth 101. Treatment of Age-Related Macular plasminogen activator and factor exerts anti-inflammatory D e g e n e r a t i o n w i t h receptor system in a mouse effects on human retinal Photodynamic Therapy (TAP) model. Arch. Ophthalmol. 2004; pigment epithelial cells. A Study Group. Verteporfin 122: 1844–1849. potentia l mechanism for therapy of subfoveal choroidal

leukocyte compartmentali-neovascularization in patients 112. Miyamoto K, Khosrof S, Bursell S E. et with age-related macular al. Vascular endothelial growth zation. Invest Ophthalmol Vis d e g e n e r a t i o n . A r c h . factor (VEGF)-induced retinal Sci 2003;44:3947.

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A SNMINI REVIEW 72


B. W. Gawali , P. B. Rokade , G. B. Janvale and S. C. Mehrotra 1 2 1 1

1 2Department of Computer Science and IT, Dr. B. A. M. University, Aurangabad, Maharashtra, Balbhim college, Beed 431122 INDIA

ABSTRACT

Many clinical studies of biological signals reveal correlation between the effect of estrogen, progesterone and brain signals captured in the form of Alpha, Beta, Theta and Delta by Electro Encephalography (EEG) recordings. It is known that there is significant and sustained changes in the levels of sex hormones that may have profound effect on brain functioning leading to changes in mood, memory and learning in women. The theta oscillations are involved with the memory encoding processes and anxiety; the alpha oscillations are associated with memory processing, whereas beta and delta pertain to attention. The release of the estrogen and progesterone hormones is divided into three phases that is premenstrual, menstrual and postmenstrual states. The frequency patterns of the brain signal and hormones are reported to be correlated. We have reviewed the use of EEG recordings in interpreting the variations in estrogen and progesterone level with the purpose of highlighting the area for further research. This technique can also be applied to study other biological signals, their frequencies and relative phases.

KEY WORDS : Alpha, Beta, Theta, Delta, EEG, Estrogen and Progesterone.

Corresponding Author : Bharti W. Gawali, Aurangabad-431002, (Maharashtra State) India. Mobile : 9422552360 [email protected]

Introduction Physiological Effects of Estrogen and Effect of estrogen and progesterone on Progesterone brain The electroencephalogram (EEG) is a In order to understand the role of Several studies indicate that there is a recording of the electrical activity of the estrogens and progesterone it is brain. The first recordings were made by relationship between ovarian hormones important to remember that hormones Hans Berger in 1929 although similar and functioning of central nervous

studies had been carried out in animals as are vital chemical substances often system. The amount of these hormones early as 1870. The waveforms recorded referred to as "chemical messengers,"; produced by the body can vary from are thought to reflect the activity of the which carry instructions from one group month to month and year to year surface of the brain, the cortex. This of cells to another. Hormones regulate the depending on many factors including activity is influenced by the electrical growth, development, sexual function stress, nutrition and exercise. Ovarian activity from the brain structures and moods. Estrogen and progesterone steroid hormones affect neurotrans-underneath the cortex. are such hormones. In women, estrogen is mission in the brain. Recently, paired Tran

circulated through the bloodstream and it cranial magnetic stimulation (TMS) was is binds to estrogen receptors on cells in used to measure the effects of the targeted tissues, that affects the breast, 3menstrual cycle in normal women . The uterus, bone, liver, heart, and brain. effect of ovarian hormones during the Progesterone hormone is secreted by the

menstrual cycle has been reported to ovaries. It stimulates and regulates various

influence several aspects of cognitive and functions. It plays a very important

motor functions in women including function in maintaining pregnancy and

spatial and verbal abilities, visual memory, helps prepare the body for pregnancy by

and motor performance. Menstrual regulating the monthly menstrual cycle. phase-dependent changes in emotional The pituitary gland in the brain generates processing as well as in mood and anxiety hormones follicle-stimulating hormone Figure 1 : 10/20 System of electrode have also been reported. Indeed, while [FSH] and luteinizing hormone [LH]. In placement everyone thinks of hormones as the reproductive years, the release of these chemicals that drive our reproductive Small electrodes are placed on the scalp in hormones causes the new egg to mature system, in truth, there are receptors for special positions. These positions are and be released from its ovarian follicle both estrogen and progesterone through-identified by the International 10/20 each month. As the follicle develops, it

System. Each electrode site is labeled with out our body. When these hormone levels produces the sex hormones called a letter and a number. The letter refers to begin to decline, as they do in the months estrogen and progesterone. The the area of brain underlying the electrode and years leading up to menopause, every physiological effects of the estrogen e.g. F - Frontal lobe and T - Temporal lobe. system that has these hormone receptors leads to various symptoms like increased Even numbers denote the right side of the registers the change including brain. A body fat, weight gain, depression, anxiety, head and odd numbers the left side of the disruption in an entire chain of bio-headaches where as progesterone leads

1head . 10/20 system of electrode is shown to sleepiness, depression and prevents chemical activity, which in turn affects the 2in figure 1. cyclical migraines respectively . production of mood-regulating chemi-

Ovarian hormones and the brain signals

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A SNMINI REVIEW

cals, includes serotonin and endorphins. National Academy of Sciences, shows consequently sampled. The absolute The result of these changes are mood contrasts in activity over the course of a power of the brain signal was then noted. swings and depression. month and provides a baseline for The study showed the following

understanding the emotional and observation:In the study, carried out by Jen-Chuen behavioral changes that 75 percent of all Hsieh and colleagues, they identified the ? In premenstrual period progesterone women report experiencing before, region of the brain used for coping with is relatively higher as compared to dur ing and af te r the i r per iod . stress, flips to the opposite side of the estrogen and alpha signal seems to Protopopescu and his team used brain during a woman's menstrual cycle be dominant followed by the delta functional magnetic resonance imaging from an area linked to negative emotion and yheta.(FMRI) to monitor the activity of the to one that usually deals with positive ? I n m e n s t r u a l p e r i o d t h e orbital frontal cortex, which is known to thoughts. The researchers studied 14 progesterone is considerably high be associated with regulating emotion women using a magneto encephalograph and Delta seems to be dominant and controlling behavior. To capture the -a machine that measures magnetic waves followed by Alpha and Theta.activity, 12 healthy women in the age created by brain activity.

? In Post menstrual, the progesterone group of 22 to 35 were considered. The All the subjects were right-handed, to goes high and estrogen is relatively subjects were experimented with a series ensure that the left-right orientation of low and Alpha is prominent followed of negative, neutral and positive words their brains matched. When the subjects by Delta and Theta.meant to illicit emotional responses. The were shown frightening images, there subjects were tested before and after their Brain signals have been extensively was activity in the right half of the menstrual cycle. They result of experiment studied and associated in literature, with subject's brains. This side of the brain 13-15is reported during the one to five days functional abilities . Theta oscillations tends to process negative feelings, such as before menses and the subjects showed in human beings are involved with anxiety. However, during the women's greater activity in the middle front part of perceptual and memory encoding menstrual periods, the images activated the brain region and less activity on the 16processing, verbal and nonverbal task areas in the left half of their brains, which sides. After menses, more activity 17and it is also related to women anxiety . handles positive emotions. The switch in occurred on the sides than the middle Alpha oscillations have been observed in brain dominance during menstruation front area. 18memory processes and Delta and Beta could help women cope with stress linked

19According to the report, the reallocation 4 oscillations is observed in attention . to hormone changes . of activity from one part of the brain Several studies have also reported the

The other study used invivo- functional region to another may reflect the organ's possible relation of the different brain neuroimaging providing the means to ability to compensate for hormonal signal with the sex hormones.probe brain activity in a noninvasive changes and help a woman maintain a

Conclusionmanner and is well suited to monitor consistent emotional state. The scientists The working of brain is a function of the changes in emotional brain activity as a are now working to compare these results status of ovarian hormones. The function of menstrual cycle status. Studies with imaging work on subjects that disturbances, lack of sleep and many that used positron emission tomography experience more severe premenstrual

6 symptoms of menopause and various (PET) have provided evidence that the mood symptoms .other emotional state modify women's menstrual cycle influences resting brain Sex hormone levels and brain waves are behavior making it difficult to analyse.glucose metabolism and patterns of brain biological signals of interest in many areas

activity during cognitive and emotional of science. Research is been carried out Referencesprocessing. The experiment was carried using FFt and EEG profiles to get the 1. Saeid Sanei J A “EEG Signal Processing“ Wiley out by showing the printed words with Publication, 2007.correlation between the brain signals – negative, neutral or positive pictures. The 2. Wright K P, Badia P. “Effects of menstrual cycle Delta, Theta, Alpha, Beta and estrogen blood oxygen level is recorded in their phase and oral contraceptives on and progesterone in women. There is

alertness, cognitive performance, and brains, which corresponded to increased evidence that in healthy young women circadian rhythms during sleep. brain region usage. The data was these signals and other ones are Deprivation. 1999; 103: 185-94.

segregated between premenstrual and 7 - 10correlated . The absolute power of EEG 3. Smith M J, Keel J C, Greenberg, et al. postmenstrual situations. Premenstrual profile of women with normal menstrual ”Menstrual cycle effects on cortical patterns help to control the emotion 11, 12 excitability” American Academy of cycle and variation in estrogen and

Neurology 1999.where as reciprocal effect is seen in progesterone have also been previously 5 4. ht tp : / /www.med ind ia .ne t /news /P re -postmenstrual phase . studied. Some of these studies included

menstrual-Stress-Makes-the-Female-For the first time, scientists have brain waves that were sampled twice in Brain-Take-U-turns-44356-1.html.pinpointed an area of the brain involved in each period corresponding to the 5. http://www.sciam.com/article.cfm?id=brain-a woman's menstrual cycle. The research, following days of menstrual cycle. The images-reveal-menst.

reported online by the Proceedings of the progesterone and estrogen levels were 6. Becker D, Creutzfeldt O D, Schwibbe M, et al.

A SNMINI REVIEW 74


Changes in physiological, EEG and psychological anxiety along the menstrual cycle”, International Journal of Phychophysiol parameters in women during the Revista Mexicana De Psicologia 2002; 1996; 171 : 161-171. spontaneous menstruation cycle and 187-195.

17. Fernandez G, Griff A, Vonoertzen J, et al. following oral contracep-tives, 12. Zani, Proverbio A M, in the Cognitive Menstrual cycle dependent neural Psychoneuro-endocrinology 1982 ; 75- electrophysiology of mind and

90. plasticity in the adult human brain is brain academic Press, san Diego CA , hormone, task and region specific, 7. Majewska M D, Steroids and brain activity, USA 2003; 377-400.

essential dialogue between body and journal of Neuroscience 2003; 23: 13. Steriade M, “Grouping of brain rhythms in mind, Biochemical pharmacology corticothalamic systems,” journal of 3790-3795.1987; 36 : 3781-3788. Neurosciences 137 , 1087-1106, 2006.

18. Hollander, Hausmann M, Hamm J P, et al. Sex 8. Smith M J, Adams L F, Schmith P J, et al. “Effect 14. Hwang G, Jacobs J, Geller A, et al. EEG hormonal modulation of hemispheric of ovarian hormones on human cortical correlates of verbal and nonverbal

asymmetries in the attentional blink, excitability, Annals of Neurology 2000; working memory, Behavioral and brain 51 : 599-603. Journal of International neuropsyco-Functions 2005; 1-20.

logy soc. 2005; 11: 263-272. 9. Walpurger V, Peitrowsky R, Kirschbaum C, et 15. Tesche C, Karhu J, “Theta oscillations index al. Effect of the menstrual cycle on human hippocampal activation during 19. Corsi-Cabrera M, Galindo-Vilchis L, Del Rio auditory event-related potentials, a working memory task, Proceeding of Portilla Y, et al. Within-subjects Hormone Behavior 2004; 600-606. National Academy of Sciences USA

reliability and inter-session stability of 10. Solis-Ortiz S, Corsi-Cabrera M, Ramos J, et al. 2000; 97.

EEG power and coherent activity in EEG oscillations during menstruation 16. T. Harmony, Femandez T, Silva J, et al. “EEG women evaluated monthly over nine cycle, International Journal of Neuro- Delta activity: An Indicator of attention

science 1984; 279-292. months, Clinical Neurophysiology to internal processing during 11. Solis S, Corsi-Cabera M, “EEG pattern of performance of mental tasks. 2007; 118: 9-21.

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The International Brain Research Organisation (IBRO) recently advances in molecular biology and whole genome approaches organized a two week summer school on Neural Degeneration have identified a number of gene mutations that appear to be and Regeneration in the University of British Columbia(UBC), predisposing factors for protein misfolding and aggregation in Vancouver. Twelve students from India, Argentina, Brazil, such disorders. Accumulation of aggregated proteins inside Indonesia, China, Morocco, Kenya and Cuba, were selected from neurons or in the neuropil leads to synaptic dysfunction, neuritic some 130 candidates to travel to UBC and interact with some of alterations, neuronal death, and disease symptoms. The future the leading group leaders in the campus and share their therapeutic interventions in neurodegenerative diseases might thoughts through a unique inhouse interactive program. The include the reduction in apoptosis, protection and plasticity, the initiative of IBRO seeks to build capacities in the developing molecular modification of aberrant genes, and artificial countries by picking budding researchers from these countries induction of neurogenesis.such that the trained individuals are able to positively transform The bioinformatics training session was very useful. Several the scientific environment of their home countries when they

neuroscientists with expertise in genetics, molecular and cellular return back. In a marked departure from sponsoring such

biology, transgenic mouse modeling and informatics, focused scholars for attending big meetings, such small groups witness

on developing tools to understand the molecular mechanisms of rdenhanced perspective development. The school followed the 3

neurodegenerative disorders by combining a multidisciplinary Canadian Association of Neuroscience meeting were all students

approach. It was particularly interesting to attend sessions by presented their posters and had a chance to interact with the

clinician-scientists and neuroscientists who emphasized the Canadian neuroscience community at large.

power of collaboration in advancing our knowledge about The school included talks (presentations, lectures) from leading neurodegenerative diseases. The neuroethical implications of researchers and clinicians who discussed the pathological and the application of these technologies in understanding the brain medical sequelae of various neurodegenerative diseases were also introduced. The daily sessions finished with a general including the approaches that promote regeneration and repair. discussion where the interaction between the faculty and the The students were encouraged to be active participants, not only students led to useful leads and interesting debate. Majority of to ask questions but also to present their own research. The such sessions lasted until late night.sessions were arranged in various centres in the campus which

The role of immigrants in the development of science in Canada exposed the students to the strong infrastructure facilities

and the world was also emphasised. A number of discussion-enjoyed by UBC researchers. The school was hosted at University

intensive sessions were devoted to elements of scientific career of British Columbia- (UBC) affiliated research centres such as

development including discrimination and bias (racism, sexism, Michael Smith Labs, Life Sciences Institute, Brain Research

age-ism). Strategies for developing mentorship networks in Centre, Centre for Molecular Medicine and Therapeutics and

order to maintain and enhance career development were iCORD. The sessions were designed in such a manner that these

discussed periodically. This school was preceded by the meeting evoked interest in linking the basic research to translational

of the Canadian Neuroscience Society which provided a unique research with possible therepeutic interventions. Students were

opportunity for these students to interact with Canadian encouraged to consider humanistic and ethical principles in

researchers. The students were encouraged to put up their performing their research by exposing them to special session on

posters at this meeting. Majority of these students attending the neuroethics.

course chose to stay for an additional 1-2 days at the end of the The neurodegenerative diseases of the nervous system reduce formal program which showed the linkages they had developed the quality of life of millions of people worldwide. to the program mentors and other labs. All students were Neurodegenerative diseases like Alzheimer's, Parkinson's disease provided with course content and a resident accomodation. and amyotrophic lateral sclerosis are considered one of the Their stay was moderated by the recruitment of local trainees major challenges of modern medicine. In the past several which acted as their hosts and mentors.decades enormous effort has been placed into resolving the

Facultymechanism(s) of neurodegeneration but in spite of great UBC: Jane Roskams, PhD, Peter Rieckmann, MD PhD, Jon Stoessl, advances many areas remain to be explored as lucidily presented

by one of the pioneers of the field – Prof. Patrick McGuire of UBC. MD, Wolfram Tetzlaff, MD, PhD, Blair Leavitt, MD PhD, Lynn Majority of these advances were discussed in the invited talks. Raymond, MD, PhD, Martin McKepwn, MD, PhD, Tim Murphy, Thus some presenters argued that protein misfolding, PhD, Yu Tian Wang, PhD, Anthony Phillips, PhD, Judy Illes, PhD, aggregation, and accumulation represent the key pathogenic Brian MacVicar, PhD, Dan Goldowitz, PhD, Kurt Haas, PhD, events common to most neurodegenerative diseases. The Shernaz Bamji, PhD, Joanne Weinberg, PhD, Catherine Rankin,

3rd Canadian IBRO school of neuroscience :neurodegeneration and regeneration

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A SNREPORT

PhD, Tania Lam, PhD, Christopher Loewen, PhD, Tim O'Connor, and imaging software; (5) Evening ethics discussions on the role PhD. McGill University and IBRO Ante Padjen, MD, PhD of international scientists on the design and implementation of

science and health policy, the reporting of scientific fraud, the Trainee Faculty (MD, PhD students): Will Guest (ALS), Andre regulation of clinical and pre-clinical “trials” and the role of a Gaudet (Peripheral nerve regeneration).support network in helping to maintain the optimal mental

Outside UBC: Sharon Louis and Allan Eaves (Stem Cell health and well being of developing scientists. Students actively

technologies), Mike Hawrylrycz (Allen Institute for Brain participated in all the school activities: Theoretical classes

Research)(interesting questions and discussions), Practical Activities,

Summary (1) Dr Sir John Sulston (Nobel laureate, 2002) Students presentations and Round Tables were as interesting as presented a lunchtime talk and discussion on the role of ethics beach volleyball and soccer. All students and some tutors were and intellectual property in advancing science and medicine in lodged in Green College on the BC campus. All lectures were developing countries. (2) Dr Pat McGeer gave an engaging and video recorded as part of the IBRO Edu learning resources and informative after dinner presentation on the history of will be made available on the web shortly.neurodegenerative disease, and hopes for the future. (3) Students completed a visit to a biotechnology company, Stem Akshay AnandCell Technologies, to learn how individuals make career choices Neuroscience Research Lab, Department of Neurologyto enter industry, and to understand how biotech companies Post Graduate Institute of Medical Education and Researchwork; (4) Hands-on bioinformatics training on genomic analysis Chandigarh

*Based on the IBRO School organised at UBC, Vancouver from May 24 to June 3, 2009.

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CASE REPORT

Introduction no evidence of acute inflammation. When lower end of shunt was exteriorized, the pus drained grew acid fast bacilli and the

The widespread prevalence of tubercular meningitis among wound margins of abdominal wall when biopsied also

Afro-Asian population continues to indicate the socio-economic confirmed the granulomatous inflammation, consistent with disparity between Eastern and Western parts of world. Eighty tubercular wound or ulcer. The abdominal shunt wound gave percent of tubercular meningitis develops hydrocephalus, of way and the pus got evacuated, leaving a typical punched out 1which 65% require CSF diversion procedure . At our institution, tubercular ulcer (figure 2). Ultrasonography showed parietal 1004 patients with tubercular meningitis have undergone CSF wall pus collections, with echogenic debris. CT scan of upper diversion surgery in the last one-decade. Two-third of patients abdomen revealed pus pocket with in parietal wall of upper belongs to pediatric age group and twice is males. We present a abdomen (figure 3). The whole shunt assembly was removed unique case of tubercular meningitis, which had dissemination and the patient continued on anti-tubercular therapy. At 6 of tubercular bacilli along the shunt, even while on anti-months of follow-up, the child is asymptomatic with completely tubercular chemotherapy. healed ulcer. The child did not require cerebrospinal fluid

Case Report diversion procedure at follow-up.

A 3-year–old female presented with altered sensorium and low-Discussiongrade fever of 6 months' duration. After investigating with CT

2The tubercular bacilli positivity is 30% in tubercular meningitis . scan of head and chest radiograph, the child was diagnosed to Our case had miliary tuberculosis, hydrocephalus, mediastinal have miliary tuberculosis, tubercular meningitis and consequent adenopathy, as well as tubercular bacilli positivity. The hydrocephalus (figure 1). The child was started on anti-extracranial dissemination of tubercular bacilli from ventricular tubercular chemotherapy, consisting of 4-drugs(Isoniazid,

Rifampicin, Pyrazinamide, Ethambutol) and a ventriculo- CSF, occurred through shunt tract. A fatal case of tubercular peritoneal shunt was placed. The CSF culture taken during shunt bacilli dissemination through ventriculo-atrial shunt, causing

3insertion was positive for tubercular bacilli after 6 weeks of recurrent miliary tuberculosis has been reported earlier . Hence, incubation. After remaining asymptomatic for 1 year, the child the prosthesis induced bacilli transmission and spread has been complained of fluctuant swelling all along the shunt tract, with documented earlier. However, the persistence of bacilli after one

Dissemination of tubercular bacilli by shunt placement

Sandeep Mohindra, Rahul Gupta and Rajesh Chhabra

ABSTRACT

A unique case of tubercular meningitis, developing shunt complication is being reported. We present a pediatric patient with tubercular ulcer formation at abdominal shunt insertion site, marking a grotesque form of tuberculosis. The final outcome of the child was satisfactory after complete course of chemotherapy.

KEY WORDS : Trigeminal autonomic cephalagias, Headache, Hypnic.

Corresponding Author

Sandeep MohindraE-mail : [email protected] : 0172-2756699, 09815535675Fax : 0172-2744401

Department of Neurosurgery, Postgraduate Institute of Medical Education and Research, Chandigarh, India

Figure 1 : Axial section of contrast CT scan of head showing tubercular hydrocephalus.

Figure 2 : Clinical photograph of abdominal wound of shunt placement, showing a typical tubercular ulcer. The ulcer is typically punched out, with vertical walls, without any signs of inflammation.

A SN 78


CASE REPORT

1sterilize the lymph nodes . The persistence of bacilli for such a 1prolonged duration makes the present case exceptional .

We propose that a minimum of 2 weeks should lapse before inserting the shunt, so as to avoid such a complication. The gold standard of management remains full dose chemotherapy for

1prolonged duration of 12-18 months .

Conclusion

Tubercular bacilli may persist, in spite of full dose of chemotherapy. A latent period of 2 weeks may be provided, before inserting shunt, so as to minimize the chance of bacilli dissemination.

References

1. Van den Bos F, Terken M, Ypma L, et al. Tuberculous meningitis and miliary tuberculosis in young children. Trop Med Int Health 2004 ; 9 : 309-313.

2. Thwaites GE, Hien T T. Tuberculous meningitis ; many year of chemotherapy is noteworthy. The alternate explanation questions, too few answers. Lancet Neurology 2005 ; 4 is the spillage of tubercular node content into the shunt tract, : 160-170.making a subcutaneous cold abscess.

3. Shibolet S, Dan M, Jedwab M, et al. Recurrent miliary After initiating chemotherapy, the time taken to sterilize the tuberculosis secondary to infected ventriculoatrial

shunt. Chest 1979 ; 76 : 328-330.ventricular CSF is usually 2 weeks, while it takes 4 weeks to

Figure 3 ; Axial section of abdominal CT scan showing a pus collection with in parietal wall, a cold abscess.

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A SNBOOK REVIEW

4 of section 2 the author has attempted to deal with these to certain extent. For cell cultivation, proper design of culture medium is very important. It plays a vital role for considerations on medium selection, medium constituents taking care of proximate and ultimate principals in which various supplements need to be given first priority. This information has been nicely organised in Chapter 5. In Chapters 6 and 7 the author deals with the needs to begin a productive animal cell culture facility, efficient laboratory design and judicious selection of equipment and infrastructural requirements in monitoring and regulation of environmental parameters for maximization of product formation.

In Chapter 8 the advantages of a single cell type system for which separation of viable cells of pure/ mono culture is of importance has been explained. Separation based on physical and surface

Animal Cell Technology : Authored by Asok Mukhopadhyay, properties of cells are also presented. Chapter 9 of the book

published by I.K. International Publishing House Pvt. Ltd., Green brings out the necessity and importance of quantitative analysis

Park Extension, New Delhi-110016, pp 380, price Rs 795/- .of each experiment and interpretation of results. For this various

Present century is a century of cells. Animal cell biology is being equipment are used in analyzing culture medium thoroughly. exploited extensively considering needs of various aspects of Chapter 10 of Section 3 describes that since each single animal animal cell technology. This book 'Animal Cell Technology' is cell acts as a production factory for important utilisable highly specialized treatise. Although other books have appeared product(s) the most crucial need is to stabilise the cell in terms of earlier in international market with similar titles, the contents are stable and high level gene expression of target product/protein. different. This research based book, in my opinion, is intended Chapter 11 and 12 of the same section describe the importance for readers who study techniques involving animal cell culture of cellular engineering considerations for production of inorder to deliver products of today and have prospects for years therapeutics from laboratory scale to large scale in a to come. The book has been packed with updated research manufacturing process. In transferring laboratory scale data to findings on animal cell technology. Since the author has a long large scale production study on scale up criterion in bio reactors association with animal cells research area he is certainly well is important in terms of transport phenomenon involving animal qualified to undertake the difficult task of producing such a nice cell culture bio processing. For facilitating transport book. The entire book has been divided into four sections phenomenon in terms of oxygen mass transfer to respiring cells comprising of a total of 16 chapters. what type of bio reactor is most suitable chapter 12 deals with

the same. Chapter 1 in Section 1 introduces readers to organizational/ structural design features and special properties of mammalian Chapter 13 of Section 4 provides that based upon the cells. In Chapter 2 the author presents molecular mechanism knowledge of previous sections, development of several involved in animal cell proliferation and cell death. In describing advanced cell culture technologies have occurred and many are this he has first dealt with animal cell growth phases involved in yet to come up. As for example embryonic and adult stem cells, cell cycle and later how growth regulation is achieved. In this cloning and gene therapy, regenerative medicines are few to regulation the mechanisms of cell signalling is also described. name. Readers of the book may learn their importance. Also Animal cells have proliferation, differentiation and death present trend and future perspectives as presented by the phases. Death of mammalian cells has been stated to occur in author, one may feel the importance of animal cell technology in two ways; necrosis and apoptosis by programmed manner. sophistication of life. Inclusion of a summary Para at the end of Means of detecting necrotic cell death (NCD) and programmed each chapter of all sections is helpful to understand the content cell death (PCD) and inducing factors are dealt in this chapter in brief. Finally, the pages of Glossary and Index in the book will effectively. help the readers of the book.

In Chapter 3 of the same section the reader may notice This book, in my opinion, will be a welcome addition to many differences between primary cells, cell strain and cell line. These bookshelves, libraries and the author needs to be congratulated cells may be used for culture development in manufacturing for bringing out this up to date research based book.biological products of industrial/commercial importance. For

S.N Mukhopadhyaymanufacturing animal cell products the knowledge of preservation of material is required which includes issues of Professor, D.B.E.Bpurity in basic cell culture technology and hardwares. In Chapter I.I.T, Delhi

Asok Mukhopadhyay

A complete culture handbook

Reviewed by : S.N. Mukhopadhyay

I.K. International Publishing House Pvt. Ltd.; 2009Hard Cover, Size : 6.5x24.5 cmPages : 380; Rs. 795/-ISBN : 978-8-18986-696-9

Animal Cell Technology

80A SNMOL SHOTS


Molecular Shots

Increased astroglyosis visualized by GFAP immunostaining around amyloid in cortex of 7 month old APP/PS - 1 mouse. Scale bar: 20 ?m

Gururaj Joshi and Jeffrey A Johnson, School of Pharmacy, University Wisconsin-Madison.

? plaques (arrow)

? III Tubulin GFAP Merged + Hoechst

??

Gururaj Joshi and Jeffrey A Johnson, School of Pharmacy, University Wisconsin-Madison.

III tubulin and glial fribrillary acidic protein (GFAP), immunoflorescence in primarycortical cultures from E15 mice. Scale bar: 20 m

4G8 GFAP Merged + Hoechst

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A SNMOL SHOTS

Neuronal (green) and glial cell (red) distribution in hippocampal area of 12 weeks old male mouse.

Hoon-Ki Sung and Paria Mohseni from Nagy Lab, Toronto, Canada.

nucleusglial cell (GFAP)neuron (NeuN)

Differentiation of murine embryonic stem cells into neural lineage :1. Neurons, 2. Astrocytes, 3. Oligodendrocytes

MAP2

AstrocytesNeurons

GFAP

Oligodendrocytes

04

Nibedita Lenka, NCCS, Pune

Editor in Chief

Co-editors

Akshay Anand graduated from Post Graduate Institute of Medical Education and Research (PGIMER). After obtaining his post doctoral training at Department of Immuno-pathology in the same Institute he joined the University of Kentucky as Scientist II. After serving their for two years, he returned to join faculty at Department of Neurology and Institute Stem Cell facility, PGIMER. He runs the Neuroscience research cum diagnostic facility and is the co founder of Leukemia Research Foundation. Dr Anand is a research improvement specialist who is interested in understanding the processes of retinal degeneration and regeneration using invitro, invivo and biotherapeutic approaches. His other research interests range from

analyzing the role of genetic and other risk factors that modify Stroke, PD and AMD to screening of novel drugs in memory enhancement. He has also been involved in the discovery of animal model of AMD published in Nature Medicine. Dr Anand is a recipient of Young Scientist Award from DAE (2005), Retina Research Foundation / Joseph M. and Eula C. Lawrence Award (2003), served as Judge at ISEF, USA (2002-3), obtained over a dozen International travel grants, and serves on Editorial board of Neuroepidemiology. He is also Reviews editor of Frontiers in Behavioral Neurosciences. Dr Anand is the guest faculty in the Human genomics program of Panjab University and advisor to the local chapter of Indian Muscular Dystrophy Association. He was recently designated Expert on Mission by ICGEB, Trieste for Intellectual property and is resource faculty for various national and International forums

Anand Swaroop is currently the Chief of the Neurobiology-Neurodegeneration Repair Laboratory (N-NRL) at the National Eye Institute, National Institutes of Health in Bethesda, MD, USA. He graduated from Indian Institute of Science, Bangalore, and did post-doctoral studies at Yale University. He joined University of Michigan Medical School faculty in 1990 as Assistant Professor of Ophthalmology & Visual Sciences and of Human Genetics. Before joining NIH, Dr. Swaroop held Harold F. Falls Professorship at the University of Michigan. In September 2007, Dr. Swaroop joined the National Eye Institute as Senior Investigator to

establish a new program for developing knowledge-based treatment paradigms for retinal diseases. Using genetic, molecular and biochemical approaches, the Swaroop lab focuses on the development and function of the neural retina, photoreceptor differentiation and mechanism of diseases such as retinal and macular degeneration due to hereditary and environmental factors. In recognition of his leadership, Dr. Swaroop has been the recipient of numerous honors including the Board of Director's award from The Foundation Fighting Blindness for outstanding research in 2006 and the Harrington Senior Scientific Award from Research to Prevent Blindness. In July 2007, he received the highest honor that the University of Michigan Medical School bestows on a faculty, namely the Distinguished Faculty Lectureship Award for his extraordinary research accomplishments and service to the scientific community. He has been a reviewer for NIH study sections, and numerous foundations including The Wellcome Trust grants, UK, The Foundation for Fighting Blindness, USA and Medical Research Council of Canada. He has been editorial board member of IOVS and Molecular Vision and reviewer of manuscripts for numerous journals in Genetics, Neuroscience and Vision research. Dr. Swaroop has several pathbreaking publications in journals such as Nature, Nature Genetics, AJHG, JBC, PNAS, etc.

Akshay Anand, Ph.D

Anand Swaroop, Ph.D

Executive Editor

the new team

A SNPEOPLE & VIEWS 82


Denis English, Ph.D

S. Prabhakar, M.D, D.M, FIAN

S Prabhakar graduated from MDU, Rohtak and completed his DM from Post Graduate Institute of Medical Education and Research, Chandigarh. He is currently the Prof and Head of Department of Neurology and

Professor in Charge of Research and Education Cell, PGIMER, Chandigarh. He has been Director of Studies, Vice President and President, Indian Medical Association, Chandigarh State Branch. He is the immediate Past President of the Indian Academy of Neurology and has earlier remained Vice-President, Treasurer for 9 years and Honorary Secretary for 3 years to serve Indian Academy of Neurology. Prof Prabhakar has been expert on the selection committee of Himachal Pradesh Public Services Commission, Maharishi Dayanand University Rohtak (Haryana) Dayanand Medical College, Ludhiana, A.I.I.M.S., New Delhi, and PGI, Chandigarh. He has published more than 200 papers in journals of national and international repute and has contributed 28 chapters in various text books. Dr. Prabhakar was elected as Chief Editor, Neurology India, the Official Journal of Neurological Society of India for a period of 6 years from 1997-2002. During his first half of tenure, 'Neurology India' has been indexed in Index Medicus and MEDLINE. He is also Co-editor

Senior Editor

Senior Editor

as Dr. Kenneth Brigham at Vanderbilt, Dr. English established the role of neutrophil-derived free radicals in inflammation and authored many of the first papers addressing processes

Denis English received his doctorate in that regulate free radical release from stimulated Microbiology from the University of neutrophils. Denis, as he prefers to be addressed, is widely Illinois School of Medicine As a recognized as a person who quickly addresses fundamental graduate student, Dr. Engl ish issues that escape those who assume greater tasks. While developed the leukocyte scan, a others assess therapeutic uses of stem cells, he asks if stem process--now replaced by MRI-- once cells exist and if so, how are they defined. He questions

widely used to localize areas of infection and inflammation. dogma others hold sacrosanct and in this area is often He recently co-authored a study that demonstrates the vindicated. "Where's the proof ?", he asked in a widely localization by MRI of stem cells infused for therapy. He is disseminated editorial published as others touted the currently Professor of Neurosurgery at the University of miracles of stem cell therapy. He challenged the universally South Florida. In addition, he directs the Florida Foundation accepted policy which prohibits public disclosure of results for Developmental Research, a non profit organization that prior to peer review simply by asking why this policy exists, was created to address the reasons that stem cell therapy has and he still awaits an answer. He has challenged the very not achieved the therapeutic goals anticipated long ago. In foundations of the process of peer review as flawed, stating accord, Dr. English founded the peer reviewed journal Stem that while necessary, the process often is quite effective in Cells and Development to address these issues as it perpetuating defective dogma as it precludes publication of questioned dogma that developed too rapidly in a fast alternative results, maintains a status quo that needs to moving field. Dr. English serves as a Senior Editor of Acta change and may result in theft of intellectual property. His Medica and Microvascular Research. He has served as goal is to alter current modes of scientific communication Editor in Chief of Journal of Hematotherapy and Stem while maintaining key aspects developed during the course Cell Research as well as Stem Cells and Development and of discovery by man.reviews papers for several journals on a regular basis. He presently serves on several NIH (USA) Grant Review Committees including those recently formed in response to President Obama's infusion of funds for research to facilitate the development of methodology to regenerate tissues and organs. His own research focuses on structural development, differentiation and the divergent role of matrix signaling in the regulation of cell functions driven by effectors of Rho family GTPases. He was first to identify sphingosine 1-phopsphate as the key link between wound healing and neo-vascularization in a short study that virtually created a new field of science. As a noted science watcher and historian, Dr. English created the discussion course A Whiter Shade of Pale as a unique forum to assess how scientific misconduct has uniquely impacted upon research developments, often with paradoxical advances. He was recently selected by the NIH to evaluate procedures used to address specific aspects of scientific misconduct at major Universities funded by the NIH and how these procedures should be altered. He established the first bone marrow transplant center in Western Canada at the University of Alberta's Cross Cancer Center in 1972, as he developed many of the techniques currently used for stem cell processing, including the only approved method for processing, collecting and storing stem cells from any hematopoietic tissue with quantative recovery, which he and Dr Hal E Broxmeyer later first employed for use with umbilical cord blood stem cells. Working with the late biochemist, Dr. Joe M. McCord, who discovered superoxide dismutase, as well

A SN83 PEOPLE & VIEWS


of Text Book of Neurology, by Indian authors, which has Z&die;'lch prize of the Max Planck Society (1995), the Faisal been published in January 2001. He has edited book International Prize (1997), the Alzheimer Award from the entitled “Taenia Solium Cysticercosis from Basic to Clinical University of Munich (1998), the Lennox K Black Prize from Science”. He is Editor of the Annual Report of the Thomas Jefferson University (2006), the Grand Hamdan Postgraduate Institute of Medical Education and Research, International Award (2006), and the Victoria Prize from the Chandigarh and also editor of `Reviews in Neurology', a Minister for Innovation.CME book of Indian Academy of Neurology for a period of 3 years from 2006 - 2008. He has recently been elected as President of the Neurological Society of India.

Moussa Youdim graduated from McGill University, Montreal and has been the founding chairman of Pharmacology from its inception from 1977 to 1994. He Colin Masters, graduated from was the Finkelstein Professor of Life University of Western Australia and is Sciences and Professor of Pharmacology currently the Laureate Professor,

at the Technion- Rappaport Family Faculty of Medicine, Israel. University of Melbourne and Executive He is Professor Emeritus and the Director of the Eve Topf and Director of the Mental Health National Parkinson Foundation (USA) Centers of Excellence for Research Institute of Victoria. He is a Neurodegenerative Diseases Research and Teaching at consultant with Melbourne Health and Neurosciences Technion. He holds the position of Honorary Professor and Australia, Chair of the National Health Committee and Distinguished Chair Professor at Hong Kong and PolyTechnic Deputy Chair of the Transmissible Spongiform Universities in Hong Kong, Yonsei University in Seoul , South Encephalopathies Advisory Committee of the National Korea, Qingdao University and Shanghai University in China. Health and Medical Research Council, and co-director of He is internationally renowned for his research in Parkinsons the Australian National Creutzfeldt-Jakob Disease Case disease (PD) Alzheimers disease (AD), Huntington disease Registry. His main research interests concern Alzheimer (HD) and ALS and drug development for these disorders and disease and related neurodegenerative disorders including for establishing the importance of monoamine oxidase and the prion diseases (Creutzfeldt-Jakob disease). His brain iron metabolism for brain function. His research at achievements have been recognised by the receipt of many Oxford University led to the discovery of the monoamine international awards. He is a prolific author covering oxidase B inhibitor, l-deprenyl (selegiline) as an anti Parkinson multidisciplinary aspects of the neurodegenerative drug in 1975 and at Technion the development of the second disorders. Prof Masters has been on more than two dozen generation of monoamine oxidase B inhibitor anti-Parkinson International journals including JAD, Brain Research, drug, Rasagiline (Azilect). He developed the multifunctional Experimental Neurology and Archives of Neurology. neuroprotective and neurorestorative anti-Alzheimer drug, Masters career has come full circle, from the pathological currently designated Ladostigil ( TV 3326), which is now in study of human brain disease to clinical trials, and from Phase IIa clinical studies and M30 series.He and his colleauges discovery and characterisation of the ASÂ aggregation in have now developed the S-iomer of rasagiline, Cardi Amit AD, to the development of a coherent theory on the (TVP-1022)as a cytoprotetive drug for cardiovascular diseases mechanisms of ASÂ amyloid production and its role in which is entering Phase I studies. He has published more than causation of neurodegeneration. This theory has become 800 scientific articles and edited 45 books and has been on the generally accepted paradigm for investigating the Editorial Board of 43 International scientific journals. He Alzheimer disease. The single, most characteristic, feature has received numerous major prizes, awards and honours of Masters research has been the way in which he has from Israel, U.S., England, Germany, Iran, Denmark, Holland approached gene and protein structure and function for a and Switzerland, including two Honoary Doctorate of variety of diseases caused by proteins that aggregate in the Philosophy, Honory Causa, from universities of Semmelweis adult human brain. He has used the resulting data to University (Hungary) and Pisa (Italy). From 1991 through generate a rich variety of discoveries that have had a wide 1999, he was a Fogarty International Scholar-in-Residence at impact in understanding normal and abnormal brain

function in disease and in ageing. Masters has won many the Fogarty International Center for Advanced Study in the prestigious prizes and awards, including the Potamkin Prize Human Health Sciences program of the National Institute of of the American Academy of Neurology (1990), the Max Health in Bethesda, USA. He is the Scientific founder of Varinel Planck Award of the von Humboldt Foundation (1991), the Inc and Varinel Ltd Registered in USA.

Youdim Moussa, Ph.D

Colin L. Masters, M.D Senior Editor

Senior Editor



Preeti Joshi, Ph.D

Ante L. Padjen, M.D, D.Sc

Pankaj Seth, Ph.D

Yogendra Kumar Gupta, M.D

Subramaniam Ganesh, Ph.D

Pharmacologist of India (APPI), AEB honours award by the Academy of Environmental Biology, C. L. Malhotra Prize of APPI etc He is has been the member of Project Advisory

Preeti Joshi graduated from Kumaun Committee/ Project Review Committee/ Research Council and University, Nanital and is Head of Scientific Advisory Committee of CDRI, ITRC, CFTRI, and Biophysics at National Institute of CCMB. He is Chairman scientific Advisory Committee of NIOH. Mental Health and Neurosciences, He is also chairman of National GLP Technical Committee of Bangalore. Dr Joshi is interested in DST. Member Apex committee CSIR-Ayush coordinated understanding the intra and partnership Golden triangle program. He has been the

intercellular communication in the nervous system and its Governing body member of Indira Gandhi Postgraduate modification in disease. Institute of Medical Education and Research, Patna , Executive

member of Navodaya Vidyalaya Samiti, and Governing body of CCRAS and CCRUM.

Ante Padjen was educated in Croatia (University of Zagreb, Croatia; 1966, MD; 1970, DSc), University of Pankaj Seth graduated from Central Edinburgh (Graduate Studies), NIMH Drug Research Centre (CDRI) and is a (1971 – 1975, Washington, DC). He senior scientist at National Brain has served as Professor at McGill Research Centre(NBRC) heading the

University, Department of Pharmacology & Therapeutics NeuroAIDS laboratory. He obtained his from 1976. His primary research interests include post doctoral training from Uniformed neuroscience, information technology and knowledge Services University, Bethesda where he worked in the area of translation in biomedicine with a particular interest in Neurovirology and Ischemia reperfusion injury. Pankaj also developing areas. He is active in national, international worked at National Institute of Neurological Disorders and professional and public organizations and is one of the Stroke (NINDS) with Dr. Eugene Major. He and his team has active leaders in IBRO. recently demonstrated that HIV-1 transactivating protein Tat

from clade B, prevalent in America and Europe, is far more neurotoxic to human neurons than HIV-1 clade C that infects almost 50% of the world's HIV-1/AIDS population. He has also established human fetal brain derived neural stem cell culture system at NBRC, which is a unique resource in the country. Y K Gupta received his MD from King Pankaj is a recipient of several awards at various levels of his George Medical College, Lucknow. He career including a Sustained Superior Performance Award by is the Professor and Head of Henry M. Jackson Foundation for Advancement of Military Department of Pharmacology, All Medicine, USA, Special Act and Service Award by NINDS/NIH India institute of Medical Sciences, in 2003 as well as Director's NIH Merit Award by National and h i s a rea o f in te res t i s Institutes of Health, Bethesda, USA. He has been invited Neuropharmacology. He has earlier speaker at several National and International meetings and also served as the Director of Industrial Toxicology Research has more than 35 publications. He is advisor, IQRA Centre, Lucknow and served as editor of the Journal, Biotechnology Sciences and editorial board member of Pharmacology and Indian Journal of Physiology and Journal of Neurovirology.Pharmacology. Dr. Gupta was President of the Indian

Pharmacological Society (2005-2006). Dr Gupta serves on selection committee of National institute of Education and Research and Public Service Commission, Himachal Pradesh, RRL, Jammu and many other selection committees

Subramanian Ganesh obtained his in the country. He has more than 150 publications and is graduation from Banaras Hindu recipient of INSA young scientist medal, Shakuntala University and is currently working as Amirchand Prize of ICMR, Chadrakanta Dandiya Prize, G.

Achari Oration Award of IPS, Major General S. L. Bhatia Associate Professor at the Department of Oration Award, Association of Physiologist and Biological Sciences and Bioengineering,

Senior Editor

Senior Editor

Associate Editor

Senior Editor

Associate Editor



IIT, Kanpur. His major research focus is to identify and characterize molecular players in neurodegenerative pathways by conducting genetic screens in affected

Ursula M. Gehling received her MD families and utilizing cellular/animal models for testing and degree in 1990 from the Philipps-validating the hypothesis. He is recipient of the National University Marburg, Germany, where she Bioscience Award for career development, Scopus young studied medicine and also started her scientists award in biological sciences, and was also residency training is as a Hematologist/ awarded the Alexander von Humboldt Fellowship.Oncologist. She is currently a Senior

Lecturer in Internal Medicine. She spent a two-year research fellowship at the University of Colorado Health Sciences Center, Denver, USA in 1994-1996 studying human stem cells. Since this time her research focuses on the biology of adult human CD133-positive stem and progenitor cells. She worked Kanchan graduated from Armed for more than 10 years at the University Hospital Hamburg-Forces Medical College, Pune and is Eppendorf. In 2004, she became Head of the Research currently working as Associate Laboratory of the Department of Hepatobiliary Surgery and Professor in the Department of Visceral Transplantation. In 2009, she started a new position Neurosurgery at PGIMER, Chandigarh. at the Paul-Ehrlich-Institute in Langen. Here, her work mainly He received presidents gold medal for deals with the safety of clinical trials therapies involving best outgoing student.and received his post graduation in tissues, cells, and advanced therapy medicinal products. Dr. surgery form PGIMER Chandigarh and M.Ch neurosurgery Gehling is Associate Editor of Stem Cell and Development and from SGPGIMS Lucknow..He is an excellent surgeon who member of the Editorial Board of 3 international journals and has completed more than 1000 surgeries including 300 has a patent on blood products from mesenchymal stem cells.aneurysms and 30 CP angle/petroclival lsesions. He

possesses versatile scientific temperament with several publications in International journals and is resource faculty for several national and international forums that include interdisciplinary deliberations.

Dr Shashi Bala Singh is the only lady Director in Defence Research and Development Organisation (DRDO) in the country and is presently heading the Defence Institute of High Altitude Vaijayanti Kale graduated from the Research (DIHAR), Leh-Ladakh, India University of Mumbai through Tata

since September 2007. She is interested in addressing Cancer Research Center (now hypophagia and cognitive function impairment at high ACTREC), Parel, Mumbai. She is altitude. Her work has led to the elucidation of some basic currently working as a Senior Scientist mechanisms involved in hypophagia at high altitude in terms at NCCS, Pune and is running a stem of taste receptor sensitivity changes. She is the fellow of cell research laboratory. She is interested in the regulation Indian Association of Biomedical Scientist (FIABMS) and of stem cell fate through the microenvironment-mediated Indian Academy of Neurosciences (IAN) and has over 60 signaling processes. Dr. Kale has been appointed as an publications apart from receiving several international and Associate Editor for the international journal Stem Cell and national journals. Shashi Bala Singh is the recipient of Dr. J.N. Development and is also appointed as SAC member for Maitra Memorial Oration award in 2003, Bharat Nirman PGIMER for stem cell research. She has filed patent Talented Ladies award in 1995, DRDO Laboratory Scientist of applications for the creation of artificial bone marrow the year award in 2006, Surg Rear Admiral M S Malhotra Prize environment (ABME) and uses thereof and also for the in 2007 and Prof. Baldev Singh Oration award in 2007. She has preservation strategies for stem cells using mannose-also received prestigious DRDO Technology Spin Off Award -binding lectins of plant origin. Dr Kale has several 2007 from Prime Minister of India in May 2008.publications in the field of stem cells.

Ursula M. Gehling, M.D

Kanchan K Mukherjee, M.Ch

Shashi Bala Singh, Ph.D

Vaijayanti P. Kale, Ph.D

Associate Editor

Associate Editor

Associate Editor

Associate Editor



Editorial Advisory Members

Zhen-Ge Luo, Ph.D

Bimla Nehru, Ph.D

Nihar Ranjan, Ph.D

Nibedita Lenka, Ph.D

R K Vasishta, MD, FRCP

Dr Vasishta has a total experience of more than 27 years in this field and has more than 120 publications and undertaken important academic responsibilities by serving through several scientific societies. His areas of special interest include

Zhen-Ge Luo graduated from Chinese Neuropathology and bone pathology. Dr conducts reporting, Academy of Military Medical Sciences teaching and research along with other administrative and (CAMMS), Beijing, China and is a consultative assignments at PGIMER.Pr inc ipal Invest igator of the laboratory of synaptic signaling, Institute of Neuroscience, Chinese Academy of Sciences. He earlier served

Bimla Nehru graduated from PGIMER, as Postdoctoral fellow, Department of Chandigarh and is currently Professor in Neurobiology, University of Alabama at Birmingham. His the Department of Biophysics at Panjab major research interests are the molecular mechanisms University. Prof Nehru has wide research underlying neuronal morphogenesis and synapse experience in the areas of metal induced formation, two basic and consecutive steps in neural neurotoxicity, aging, epilepsy, cognitive development. The long-term goal of these studies is to dysfunction, Parkinson disease and identify the targets for treatment of neurological disorders

Huntington's disease. Professor Nehru extensively explored including spinal cord injury, neuromuscular disorders, and the neuroprotective mechanism of different lead compound/ neurodegenerative diseases. Zhen-Ge has received the herbal phytochemicals (curcumin, piperine as well as prestigious China Youth Science and Technology Prize in Centrophenoxine). Particularly, her work on aluminum as well 2007. He is consultant for Trends in Biochemical Sciences, as lead toxicity has been well appreciated in the scientific PNAS, Neuroscience, Developmental Neurobiology, PLOS community which is evident from her highly cited One, B iochemica l and B iophys ica l Research publications. Prof Nehru served as the chairperson of Communications etc and has several highly cited biophysics department (1999-2001) and has won numerous publications in Nature Cell Biology, Neuron, PNAS etc.awards. Apart from neurons the focus is now being diverted to the glial cells (the secondary brain cells) in the management of oxidative stress condition as brain is more susceptible to it, especially with regards to the ex-pression of heat shock

Nihar Ranjan Jana obtained his PhD protein, HSP.from Visva-Bharati University, Shantiniketan and post doctoral training from RIKEN Brain Science Institute, Japan. He is presently

Nibedita Lenka graduated from Osmania working as Associate Professor in the University, Hyderabad holding research National Brain Research Centre, fellowships under the program of CSIR Manesar. He is a fellow of National Academy of Sciences, and is currently serving as a faculty India and recipient of DBT's National Bioscience Award for scientist at National Centre for Cell Career Development. His research centers on role of Science (NCCS), Pune. Nibedita obtained ubiquitin proteasome system in neurodegenerative and her post docotoral training from neurodevelopmental disorders.

University of Pennsylvania, USA and Institute of Neuro-physiology, Univ of Koln, Germany. Her research interests include Stem Cells and Development with special focus on Embryonic Stem Cells maintenance and their differentiation

R K Vasista graduated from Post into various lineages and the guiding cues underlying the Graduate Institute of Medical same. She has served as a member of Faculty selection Education and Research (PGIMER), committee, All India Institute of Medical Sciences and was a Chandigarh and is currently working visiting scientist under Exploratory cooperative program as Professor of Histopathology. He between Japan Society for Promotion of Science, Japan and was trained in UK in Regional DST, India. She has got several honours and awards and has a Neuropathology Centre of Northern number of cited publications and a patent to her credit. She Ireland at Royal Victoria Hospital. reviews major stem cell articles from reputed journals and also



grant applications submitted by investigators to various organs, such as, bone marrow, liver, retina etc. Dr funding agencies. Mukopadhyay has earlier been instrumental in developing the

technology for production of anti-leprosy vaccine leading to patents and highly cited papers. He is on several committees that focus on translational programs in the country and is recipient of Clayton Foundation for Research postdoctoral

Rakesh Biswas graduated from Post Research award at the time of his post doctoral work. His work Graduate Institute of Medical has been adjudged as the among the best in NII that led to a Education and Research, Chandigarh product. and is currently Professor, Department of Medicine, People's College of Medical Sciences, Bhopal, India. His interest in neuroscience is related to

his special interest in creating a 'user driven health care' Fakhrul Islam graduated from Brain community of patients and health professional users Research Centre, Aligarh Muslim (including neuroscience professionals) for better health University and is currently working in the care outcomes. Dr Biswas is the regional editor: Journal of Department of Toxicology, Jamia Evaluation in Clinical Practice, Wiley Blackwell,UK and Hamdard as Associate Professor and editor:User-Driven Healthcare and Narrative Medicine and training Neurotoxicology and Forensic reviewer for Research evaluation panel of National Digital Toxicology to the graduate students. He Research Centre, Ireland and reviewer for the journal,

was IBRO Post doctoral fellow to work at the Centre de Medical Education, UK.

Neurochemie du CNRS at Strasbourg Cedex, France from October 1985 to January 1987. He has supervised 17 Ph.D. scholars and all of whom are well placed in accomplished labs or running their own labs. Dr. Islam has published 70 research papers in international journals of repute and has completed 9 ambitious projects. The main goal of Islam's lab is to pioneer

Nusrat Shafiq graduated from Post treatment of Neurodegenerative diseases. i.e. Cerebral Graduate Instiute of Medical

ischemia, Parkinson's & Alzheimer's diseases using herbal Education and Research, Chandigarh

extracts and active principles of medicinal plants and and obtained her DM from the same

nanodrug delivery.Institute. She is currently working as Assistant Professor at the Department of Pharmacology and has more than

35 indexed publications to her credit. Dr Nusrat has organized GCP workshop in her Department as its Joint Paramjeet Singh graduated from Post Secretary. Graduate Institute of Medical Education

and Research, Chandigarh earning his MD in Radiodiagnosis. He is currently working as Additional Professor and possesses 20 years of experience. He is interested in Magnetic Resonance Asok Mukhopadhyay graduated from

Imaging applications particularly in Neuroradiology, IIT, Delhi and is currently the Senior Musculoskeletal Radiology and body applications. He is a Scientist with National Institute of member of several learned bodies and has certificate of Immunology(NII), running a produc-completion of specialist training (CCST) in Clinical Radiology tive stem cell facility. He received his from the Specialist Training Authority (STA) of the Medical post doctoral training at MD Royal Colleges, UK and benn Visiting Professor at Division of Anderson Cancer Center and also served at Thapar Diagnostic and Interventional Neuroradiology, Department of Corporate R & D Center (TCRDC). He is trying to develop Radiology, University of Rochester Medical Center, Rochester, bioreactor for expansion of engraftable stem cells and also NY, USA. May, 2005. He has more than 80 publications and interested in understanding in vivo trafficking and grafting

of normal and ex vivo manipulated stem cells in different several chapters in books.

Rakesh Biswas, M.D

Fakhrul Islam, Ph.D

Nusrat Shafiq, M.D, D.M

Paramjeet Singh, MD

Asok Mukhopadhyay, M.Tech, Ph.D



Almaz Aldashev, Ph.D G.U. Gurudutta, Ph.D

Stefan Glck, M.D, Ph.D

Roberto Agustin Prado Alcala, Ph.D

Almaz Aldashev graduated from Gurudutta graduated from University of Biological Sciences and Biochemistry, Delhi and is currently the Head of Stem Russ ian Cardiological Center, Cell and Gene therapy Research Moscow, Russia, and is currently the Program, Divis ion of Radiation Director of the Institute of Molecular Biosciences. INMAS, DRDO, Delhi. He Biology and Medicine, Fellow and obtained his post doctoral training from Chief Academician Secretary of University of Sao Paulo. Brazil, University

National Academy of Sciences of Kyrgyzstan, Vice- of Pennsylvania. Philadelphia, USA and National Space chairman of Biochemical Society of Kyrgyzstan, Head of the Laboratory, Brookhaven National Laboratory, New York. Laboratory of Molecular and Cell Biology, National Center Gurudutta has interest in understanding identifying and of Cardiology and Internal Medicine. He is the Editorial validating signaling pathways, their synergism, target genes Board member of the famous national journal Izvestiya, of different germ line origin including all adult stem cels Fellow of World Innovation Foundation, Senior Fellow of (Hematopoietic, Mesenchymal etc) and Embryonic stem cells Pulmonary Vascular Research Institute, London, UK. Almaz to designate them as universal molecular manipulatable Aldashev is the author of more than 140 scientific articles, targets to enhance and harness potentials of stem cells in including 3 monographs. Almaz is recipient of Fogarty human organ transplantation & tissue engineering International Center award, British Royal Society award, applications. He is member of American society of Wellcome Trust grant, British Heart Foundation award, Biochemistry & Molecular Biology, Gene Therapy and ICSSR. Pfizer Research grant, International Sciences and USA. He is Official member of Program Advisory committee of Technology Center grant and NATO science for peace DST and recipient of Lab technology development group research award. award from DRDO. Min. of Defence. Gurudutta has been

presently exploring the feasibility of enhancement of proliferation, differentiation, survival potential and stromal-ligand adherent potential necessary for enhanced engraftment / homing of transfused quiescent CD34 stem cells by introducing genetically engineered highly potent

Stefan Glck obtained his medical genes. This strategy may possibly help to repopulate or re-degree from Free University of West generate the human bone marrow when deficiency in stem Berlin, Germany and obtained his PhD cells occurs or stem cells for transfusion are unavailable. In from RWTH, Aachen. He is a medical addition, Gurudutta has been actively studying iPS cells and oncologist as well as the Associate their mechanistic role in their recruited tissue regeneration.Division Chief of Hematology/ Oncology for Clinical Affairs. Dr. Glck

most recently was honored with the merit of Sylvester Professor of Medicine, Miller School of Medicine. He has been involved in running several clinical trials and currently

Roberto Alcala obtained his Ph.D. form serves as editorial board member for several journals and Concordia University, Canada in grant reviewer for many funding agencies. He has more Psychology. He currently holds the than 150 publications and is a resource faculty for several position of Senior Researcher at the international and national forums. He has been honored Institute of Neurobiology, National with the Best Doctors in America Award since 2006 since

his arrival to the United States of America. His interest is in University of México where he was its clinical trials in breast cancer and he is a PI of 24 clinical past Chairman. He is member of the Mexican System of studies in breast cancer in Miami, and investigator in Researchers Level III and of the National Academy of Sciences. numerous scientific, translational projects. Dr Glck is a In recognition of his scientific and academic merits he has member of such prestigious professional organizations, as been awarded the Psychological Research Prize, Mexican the American Society of Clinical Oncology, the European Society of Psychology, 1984; Miguel Alemán Valdez Prize for Society for Medical Oncology, the American and European excellence and productivity in Science, 1986; National Prize of Association of Cancer Research, and the International Psychology, México, 1995; National University of Mexico Prize Association for Breast Cancer Research. in the field of Teaching of Natural Sciences, 2000. He was



Editor-in-Chief of the Mexican Journal of Psychology (1998- a company manufacturing electronic components, Bell 2003), and is member of several Editorial Boards besides an Electronics, a company manufacturing wireless sets. Dr active member of several learned societies. His scientific Sharma is associated with Commonwealth Secretariat, Asia publications include 84 articles, 44 book chapters. Centre, U.K. He has many publications to his credit in

National/International Journals of repute. Dr Sharma is a recipient of the Jacob Wolfouitz Prize award for his research paper entitled “Simultaneous Multiple Comparisons with the Worst and Best”. One of his publication “Linearization of the

Rao M Adibhatla graduated in relationship between serum sodium, potassium Chemistry from Indian Institute of concentration, their ratio and time since death in Chandigarh Science under Prof. CNR Rao and is zone of north–west India”, has been awarded “Best Research currently research Faculty at the award”.Department of Neurological Surgery, University of Wisconsin School of Medicine and Public Health, USA. He

obtained his post doctoral training from Center for Cellular Mriganka Sur, received the B. Tech. and Molecular Biology under Prof. D Balasubramanian, degree from the Indian Institute of Hyderabad, also worked at University of Iowa, University of Technology, Kanpur, and the Ph.D. Missouri, University of Minnesota, University of Kentucky degree from Vanderbilt University, as research faculty and was also awarded the Alexander

von Humboldt fellowship during 1987-88. He has more Nashville. He is the Newton Professor of than 70 publications and awarded NIH and American heart Neuroscience and Head of the Association grants. He has been invited to several Department of Brain and Cognitive International conferences and as invited guest speaker at Sciences at the Massachusetts Institute of Technology (MIT). MIT, NIH, Univ. of Miami, and Ferrer Grupo, Barcelona, Professor Sur studies the development, organization and Spain/Elder Pharmaceuticals, India; on “CDP-choline plasticity of the brain using experimental and computational mechanisms and efficacy in stroke”and is on editorial approaches. He has discovered fundamental principles by board of many important journals and reviewer on several which neurons of the cerebral cortex are wired during grants. Dr Rao is interested in signal transduction development and change dynamically in adulthood. His mechanisms in CNS trauma and integration between laboratory has developed novel technologies for visualizing inflammation, lipid metabolism and phospholipid the activity of brain cells and synapses, and proposed homeostasis in CNS injury and disorders. He is adhoc innovative strategies for treating brain disorders such as reviewer for more than 30 journals. autism. A member of the MIT faculty since 1986, he has led

the Department of Brain and Cognitive Sciences since 1997. He has received numerous awards and honors, including the Charles Judson Herrick Award, the McKnight Neuroscience Development Award, the Sigma Xi Distinguished Lectureship, the IIT Kanpur Distinguished Alumnus Award, and the Special

Suresh Sharma obtained his Ph.D. in Lectureship of the Society for Neuroscience. He has been

the area of “Ranking and Selection”, elected to the Rodin Academy, Sweden, the American

and currently working in the area of Academy of Arts and Sciences, the Third World Academy of

Biostatistics, Department of Statistics, Sciences, and the Royal Society of the UK.

Panjab University, Chandigarh. His research interest includes Biostatistics,

Ranking and selection, Statistical Inference, Estimation and testing under ordered restrictions. He has been involved

Obaid Siddiqi is a world renowned with several projects related to “Coronary Heart Disease”, scientist and fellow of several academies. “Mother and Child Health Care” and “Public Health and He was the founding Director of National Pollution Control in Asia” (Sponsored by ICMR/WHO/HEI, Centre Biological Sciences( NCBS). His lab Boston, USA) at Post Graduate Institute of Medical is trying to understand the adaptive Education and Research (PGIMER), Chandigarh. He serves

as Consultant Statistician for number of Industries in India behavior and establishing its neural like Ranbaxy, a Pharmaceutical company, Semi-conductor, correlates by modeling learning and

Rao M. Adibhatla, Ph.D

Mriganka Sur, FRS

Suresh Kumar Sharma, Ph.D

Obaid Siddiqi, Ph.D, FNASc

Statistical Consultant



memory in larva and imago. Prof Siddiqi has numerous has highly cited publications in journals including Nature, international awards and highly cited publications to his Nature Biotechnology etc and responsible for many important credit. He has served on various national and international discoveries in the field of Stem cell biology. science committees impacting on science policies in this region.

Keiji Tanaka started his research career in the primary visual cortex, and has

Lakhotia graduated from University of gradually moved to more cognitive Calcutta and is currently Professor of areas. He got Ph. D. from Faculty of Zoology and Dean, Faculty of Science Medicine, University of Tokyo and was at Banaras Hindu University. He is a one of the founding members of the fellow of Indian National Science RIKEN Brain Science Institute, where he is Academy, New Delhi, Indian Academy now acting as the Deputy Director. His research now focuses of Sciences, Bangalore and National mechanisms of visual object recognition and goal-directed

Academy of Sciences India, Allahabad and is recipient of SS behavior by using cell-recording and lesion-behavioral Bhatnagar Prize (CSIR), J C Bose Award (UGC), Sundarlal methods on nonhuman primates. He has also pursued fMRI Hora Medal (INSA), Prof. C.N.R. Rao Education Foundation on the human cortex at sub-millimeter spatial resolution. He Award for Excellence in Research at Banaras Hindu has several publications in Science and Nature. Dr Tanaka has University. He is a member of editorial boards of several held editorial positions with Science, Neuron, Journal of journals. Prof Lakhotia is an active proponent of elevating

Neuroscience, Cerebral Cortex etc and organized several the standards of Indian scientific journalism. His lab

successful Neuroscience meetings. He is recipient of focusses on developmental roles of the noncoding hsr-

prestigious IPSEN Neuronal Plasticity Prize, Tohihiko-Tokizane omega, HSP and a few other novel genes in Drosophila

Memorial Prize, and Science and Technology Prize (Ministry of using genetic, cell biological, molecular and recombinant

Education, Culture, Sports, Science and Technology-Japan).DNA approaches. Roles of these genes in modulating apoptosis and neurodegeneration in Drosophila models of human diseases and cellular effects of certain Ayurvedic formulations using fly model are being investigated.

Martin E. Schwab has been Professor of Neuroscience at the ETH Zurich and at the Institute for Brain Research at the University of Zurich. The research group

Peter Andrews gained his Ph.D. from directed by Martin E. Schwab was University of Oxford and is currently established in 1985 at the Institute for co-director of the Centre for Stem Cell Brain Research at the University of Biology at University of Sheffield. Zurich. Prof Schwab is investigating cellular and molecular Following postdoctoral research at

mechanisms of nerve fiber growth, in particular during the Institut Pasteur in Paris and the

regeneration of injured nerve fibers in the spinal cord and the Sloan Kettering Institute in New York,

brain, as well as in relation to structural plasticity in the adult he served as research scientist on the staff of the Wistar

central nervous system. The methods stem from molecular Institute of Anatomy and Biology in Philadelphia from 1978

biology, biochemistry, cell biology and neuroanatomy. This to 1992. His research focuses on the biology of pluripotent

research has led to a new paradigm in spinal cord injury human stemcells. Among his current activities he co-

research by the characterization of a novel protein of the ordinates the International Stem Cell Initiative, which is

central nervous system, namely Nogo-A, which is a strong focused upon characterising standard markers and culture inhibitor of nerve fiber outgrowth. Function blocking conditions for human ES cells. He is also the co-ordinator antibodies against Nogo-A was shown leads to functional of ESTOOLS, a European Integrated Project of 21 partners recovery and the formation of new nerve fiber connections. under the sixth framework program. He is an active Prof Schwab is on several Eropean Neuroscience committees member of an Indo UK stem cell project and serves on the and has won numerous International awards and has several editorial boards of several journals besides reviewing highly cited publications.grants from several International funding agencies. He has

Keiji Tanaka, Ph.D

S C Lakhotia, Ph.D, FNASc

Martin E. Schwab, Ph.D

Peter W Andrews, D.Phil



Walter G. Bradley, DM, FRCP Tom Isaacs

Hctor H. Garcia, MD, Ph.D

Kanikaram Satyanarayana, Ph.D

Bradley received his medical education from Oxford and is a Fellow of the Royal College of Physicians of London and Board Certified in Neurology and Psychiatry. He is a member and former Counselor of the American Neurological Association.

He is a Fellow of the American Academy of Neurology, and has been Chairman of the Scientific Program Committee, the Science Committee, and the Workforce Taskforce. He is a member of the Association of University Professors of Neurology and has been Chairman of its Bylaws Committee. Dr. Bradley has authored 24 books, more than 200 peer-reviewed articles and 100 chapters and reviews, as well as several on-line publications on healthcare reform. He is founding editor of the textbook “Neurology in Clinical Practice” that is now in its fifth edition and has become the major textbook of neurology for residents and clinicians worldwide. He founded and edited the journal “Muscle and Nerve” from 1978 to 1988, and was Editor of the Yearbook of Neurology and Neurosurgery from 1993 to 2000. He has been a member and chairman of a number of scientific advisory and editorial boards. He was organizing secretary of the Third International Congress on Muscle Disease in 1974. Dr. Bradley has an international reputation for his contributions to the fields of amyotrophic lateral sclerosis, peripheral neuropathies, inflammatory myopathies and muscular dystrophy.

Hugo Garcia graduated from the Universidad Peruana Cayetano Heredia (UPCH) in Lima, Peru, and obtained his PhD from the Johns Hopkins University Bloomberg School Satyanarayana holds a Doctorate degree of Public Health in Baltimore, USA. in biosciences from Sri Venkateswara Currently he is a Full Professor in the University, Tirupati and is currently a

Department of Microbiology at UPCH and also heads the senior Deputy Director-General and Cysticercosis Unit of the Instituto Nacional de Ciencias Chief of the Division of Publication & Neurologicas. Hugo is an acknowledged expert on Information as also Intellectual Property Neurocysticercosis and other zoonotic diseases. He has Rights Cell of the Indian Council of Medical Research (ICMR). published over 130 papers in international biomedical He has been engaged in issues relating to publications, ethics, journals amd multiple book chapters, and also serves as intellectual property rights and technology policy issues with Associate Editor, PLoS Neglected Tropical Diseases and is special reference to developing countries for over 25 years. Member of the Editorial Board, American Journal of For over 20 years he has been training young and middle level Tropical Medicine and Hygiene, Experimental Parasitology doctors on the skills of communicating science to learned and Editorial Consultant for The Lancet. journals and has also organized workshops for medical

Tom Isaacs was diagnosed with Parkinson's at the young age of 27 and since then has done everything he can to raise funds, heighten awareness and find a cure for the condition which is perceived by many as a condition affecting the elderly alone. Having

completed his highly successful 1,250 mile sponsored walk in 1999, Tom left his job as director of a London property company in April 2002 to undertake his Coastin' challenge. By April 2003, Tom had walked 4,500 miles around the British coastline, climbed the highest mountains in England, Scotland and Wales and run the Flora London Marathon, raising over 350,000. In 2004 he was runner up in the GMTV/Daily Mirror Fundraiser of the Year Award and in 2005 he was elected Charity Personality of the Year. In the same year, he co-founded The Cure Parkinson's Trust, an organisation which has gone on to raise 2.5 million and made a significant impact on the culture and urgency of the Parkinson's community and its quest for a cure. Tom has also been a Board Member of the European Parkinson's Disease Association since 2005. He also represents the interests of people with Parkinson's on DeNDRoN (the Dementias and Neurodegenerative Diseases Research Network) and is a member of the Steering Committee for the World Parkinson's Congress 2010 and Chairs its Patient Advisory Group. Tom has written a book “Shake Well Before Use” about his walk and his experiences with Parkinson's which he conveys with passion, optimism and humour. He speaks regularly to a wide ranging audiences about his condition and also often presents on television and radio giving his own perspective as someone living with Parkinson's.



journal editors. He is also an expert in publication ethics Force. He graduated from the University of Illinois with a B.S. and was the Ombudsman for the Indian Journal of in Mechanical Engineering, received his law degree from Pharmacology, first from the developing countries. He is Gonzaga University School of Law (with Honors), and has a currently the ombudsman for Neurology India. Dr masters in Intellectual Property Law from the University of Satyanarayana has been the Chairman of the Central Houston. He has written frequently on intellectual property Institutional Ethical Committees of Dr R.M.L. Hospital, New law and American constitutional law and has been published Delhi and the CCRAS, New Delhi. He is also the editor of the most recently in the Texas Tech Law Review and the University Indian Journal of Medical Research. of California at Los Angeles Entertainment Law Review

M.R.S Rao graduated from Department Pavan Duggal graduated from faculty of Biochemistry, Indian Institute of of Law at Delhi University and is Science(IIsc), Bangalore and obtained his currently working as Advocate with postdoctoral training from Baylor Supreme court of India. He is a an College of Medicine, USA after which he acclaimed academician and serves as joined back IISc. He currently directs the consultant to UNCTAD and UNESCAP,

Jawaharlal Nehru Centre for Advanced Scientific Research, Council of Europe on Cybercrime and Asian Development Bangalore as its President and has received several Bank. He has made an immense International impact on recognitions including Shanti Swarup Bhatnagar (1998, Intellectual property, Cyberlaw, E-Commerce laws and is Ranbaxy Science Foundation Award for Basic Medical Sciences interested in Business Process Outsourcing Law, Intellectual (1999), O.P. Bhasin Award for Biotechnology(2002), FICCI Property Rights and Information Technology Law, Award for R&D in Life Sciences (2004), Dr. B.R.Ambedkar Information Security Law, Defence, Biotech and Corporate Centeneray Award for Biomedical Research(2005) and TWAS Law. Pavan is the President of Cyberlaws. Net, the founder Medal oration (2008). Prof Rao is a Fellow of all the Science of the Cyberlaw Association and the Founder-President of Academies in India,and Academy of Sciences for the Cyberlaw India. He is also a member of the WIPO Developing World (TWAS), Trieste, Italy. Prof. Rao has been the Arbitration and Mediation Center Panel of Neutrals and Member and Chairman of several Scientific Advisory serves as Chairman of the Cyberlaw Committee of Committees of DBT, CSIR and DST and has also the Chairman ASSOCHAM working closely with CII and FICCI. He is a task of Research Council of CCMB Hyderabad and IICB, Kolkatta. force member of ICT Policy and Governance Working Group Prof. Rao is first Chairman of the Board of Governors of Indian of the UNICT. He is the Legal and Policy Consultant to Institute of Science Education & Research (IISER), Trivandrum. Internet Mark 2 Project, which is examining the next level of He has published more than 130 papers in international Internet. He is also fellow of Centre for Asia Pacific journals and guided 30 students for Ph.D. Degree. Prof Rao is Technology Law and Policy (CAPTEL) at Singapore and actively engaged in research on non-coding RNA in member of Panel of Arbitrators of the Regional Centre for development and differentiation, cancer biology and Arbitration, Kuala Lumpur and Asian Domain Names chromatin biology. He is also the team leader of the CSIR's Dispute Resolution Centre at Hong Kong. He has authored NMITLI Network Project on Cancer Genomics, particularly on more than six books on various aspects of the law in the last Glioblastoma.six years.

Scientific Integrity Advisor

Avadhesha Surolia, obtained his Ph.D Michael Coblenz is a patent and from the Christian Medical College, intellectual property attorney in Vellore, under the supervision of Lexington, Kentucky, U.S.A. Before Dr.Bimal K.Bachhawat and is currently attending law school he served for the Director of NII. He earlier served as seven years in the United States Air the Chairman of Molecular Biophysics

Pavan Duggal M.R.S. Rao, Ph.D

Michael Coblenz Avadhesha Surolia, Ph.D

OmbudsmanIP Consultant

IP Consultant



Unit, Indian Institute of Science during 2001-2006 and has in the discovery of the unusual quaternary structure of been visiting scientist at Massachusetts Institute of peanut agglutinin and a novel lectin fold in Jacalin as well as Technology, Boston, USA. Prof. Surolia is recipient of the structural basis of inactivation of ribosomes by gelonin-numerous National and International awards and was also all of which laid the foundation of structural biology as it the President, International Glycoconjugate Organisation stands today in the country. His original strategies for drug during 2003-2005. Prof. Surolia's pioneering contributions and DNA delivery are widely acclaimed. Surolia has recently have strongly influenced International research on structure discovered that the fatty acid synthesis pathway in human and function of lectins, orientation and dynamics of cell malarial parasite is distinct from its human host and its surface carbohydrate receptors as well as drug and interference by triclosan, a commonly used biocide, thus molecular design and their biotechnological applications. He paving the way for novel drug development for treating played a pivotal role together with his colleague M. Vijayan malaria in collaboration with Namita Surolia.




ANNALS OF NEUROSCIENCES VOLUME 16 NUMBER 1 JANUARY 2009www.neuroscienceacademy.org.in

Dear Editor :

Dear Editor:

Our discussions of my analyses of issues vs. scientific goals of stem cell research long ago led me to propose that of many candidate countries, India should be at the very center of discussions, developments, and dialogue as the field develops. I chose India because of the in depth honesty and realistic interpretations of reports submitted from new laboratories there to the journal I founded, Stem Cells and Development to be both unique and extraordinory. Of all countries, including the USA, China, Korea, Western Europe, and Asia, I was continuously impressed by the quality of stem cell research from the few laboratories in India that rose to meet this challenge. The investigators who headed these labs were soon invited to become members of my senior editorial board, particularly Dr. Vaijayanti Kale of NCCS , Dr Gurudutrta Gangenahalli of the India Ministry of Defense and Dr. Rama Verma of IIT- Madras, who trained with me in Indianapolis, Indiana (USA) a few years earlier.

I first visited India to present the Keynote to the First International Summit on Stem Cell Research, held in Chennai, and was hosted there by you and Dr. Gangenahalli of the Ministry of Defense stem cell initiative. We certainly found a lot of common ground, as I witnessed results that changed my outlook on the therapeutic promise of stem cells. On further analysis, I discovered that your government takes an active and responsible role in approving and monitoring all clinical trials; rumors that India is used because it presents a unrestricted population of patients that can be used with little monitoring are blatantly false. In addition, I was stunned by the rapid development of stem cell research initiatives in India. While I was there, I found that similarly rapid developments occurred in the development of electronic technology in India, established only a few years as a partnership with Microsoft. I have now come to learn that individuals at Microsoft and other high tech companies here are delighted at the speed and efficiency with which these projects were put into practice in India at the benefit of end users in the United States and many other countries. It seems incredible; but it happened in less than 5 years, and now high technology encompasses a major portion of the economy

International Society for Neurochemistry (ISN)/Journal of Neurochemistry; American Society for Neurochemistry/ASN Congratulations for taking reigns of the Indian Academy of Neuro; IBRO/Neuroscience. Few things such as sending the Neurosciences official journal Annals of Neurosciences as an contents of the journal articles in upcoming issues to Editor. When I first met you during December 2008 XXVI annual neuroscientists; advertising available job positions (post-doc Indian Academy of Sciences meeting at Kochi, Kerala and and faculty position) will make the journal visible also. subsequent correspondence I was very much impressed with

your commitment, enthusiasm and hard work to reinforce the Congratulations for becoming the Editor of Annals of Annals of Neurosciences by infusing new blood to take the Neurosciences and I appreciate your untiring efforts to improve journal to new heights. the quality of the journal.I agree that Indian scientists should submit their quality research Cheers,work to Indian journals to increase the impact factor. This is a

Rao Muralikrishna Adibhatla, Ph.Dsacrifice in the beginning years to strengthen the foundation of University of Wisconsin School of Medicine, USAthe journal. I have seen enthusiastic bright young researchers

who want to share their research among their peers and graciously take the positive feedback during the discussions at the December 2008 academy meeting. We need to encourage these youngsters by dedicating few pages of the journal (Journal club) to share their views and opinions on a recently published work (In neuroscience/biology journals such as Nature Medicine; Nature Neuroscience, Journal of Neuroscience, Journal of Biological Chemistry etc.). This will make them part of the neuroscience community, have fresh look in their own perspective and boost their morale. I am positive most of my young colleagues will agree with this idea and would like to contribute and share their opinions. On the same lines readers can post comments or critique on a published article- eletter to the editor (in Annals which may corroborate another independent group's work, sometimes brought up issues that reviewers might have overlooked). Happy to note the future issues will carry these sections. I do this on regular basis and immensely benefited from the responses and at times leads to collaborations.

I agree, if you intend to providing honorarium to reviewers for their contributions as far as sufficient funds are allocated for this purpose. At the same time the articles should go through rigorous reviewing which should be the top priority in order to make the journal more visible and survive in this rapid age of neuroscience. But in some special situations, facilities and infrastructure issues may limit one's abilities to meet the standards and we should keep that in mind how that will hamper the reviewing process or comments/criticism on the papers submitted by the Indian counter parts. This is by no means I am undermining the quality of Indian science, but to cover the possible scenarios. The enhanced funding from Indian government can surpass this barrier and of course good work automatically attracts endowments.

Having an international editorial board will certainly help achieving these goals. The team of experts you are announcing as international editorial board are giants in their field and we will be fortunate to have these esteemed members shape up the journal with their constant advice and suggestions.

An independent website and a publisher will be a good idea just like Society for Neuroscience/ Journal of Neuroscience;

Letters to Editor


ANNALS OF NEUROSCIENCES VOLUME 16 NUMBER 1 JANUARY 2009 www.neuroscienceacademy.org.in

of India, one of the most productive economies in the world members of your scientific community, the military and the today. government, your acceptance of input from foreign nations and

the manner wherein you reached out to them in your genuine The commitment of the Ministry of Defense as well as many quest for help, not money nor fame, but help. Devastating Biomedical Institutes in India to work with investigators from the diseases of aging perhaps can be moderated, but only a

US and other countries and to work in collaboration with FDA combined, non-selfish approach such as that you have proposed foundations as a governing principle will certainly lead to shall reach this goal. I only hope I can help.

therapeutic advances, and these I see as quickly forthcoming. Already, eminent cardiologists of Temple University have Sincerely,committed much of their time and effort to India for both

Denis English, Ph.D.,development of therapeutic research with stem cells and just as

Professor of Neurosurgeryimportantly, to bring to India techniques otherwise not

Founding Editor, Stem Cells and Developmentavailable. These men and women appreciate the opportunities

University of South Florida College of Medicinethey are afforded in India so much that they provide state of the

1909 E Mulberry, TAMPA FL 33604art care to children of Southern India at no cost. Many times,

US Authors can submit their manuscripts to:their spouses accompany them and when they do, they often arrange with United Air or other benevolent corporations a The Florida Foundation for Developmental Researchshipment of needed items for those who do not enjoy our attn Dr English, Journalismstandard of life. PO Box 1651

PaLMETTO, FL 34220I think this to be a very fair trade off, considering the ambitious Tel.: 813-355-1994nature of all involved, the restrictive yet profitable environment Fax : 941-746-4634here and the dedication I witnessed first hand between

A SNThe Journal is a free-access, multi- communications that will foster clearer figures, tables and equationsdisciplinary publication of the Indian understanding of the Neurosciences, ? The total number of words in: (i) Academy of Neurosciences that aims to generation of better diagnostic tools, the whole manuscript; (ii) the cover new advances in Neurosciences. It encourage journalism among young A b s t r a c t ; a n d ( i i i ) t h e provides a platform for papers that range scientists and development of effective Introduction.from computational and experimental cures for Neurological disorders. All

? A list of four or five keywords not work in the neurosciences to those that fit experiments described in the ANS that appearing in the title, preceded the interface between experiments and involve the use of animal or human by "Keywords:"clinic. The Journal accepts research papers subjects must have been approved by the

as research articles, brief communi- appropr iate inst i tut ional rev iew Abstractcations, reviews, commentaries, book committee and conform to accepted Research Articles and Reviews should reviews, molecular images, student's ethical standards. have an Abstract, which appears before perspectives on published reports in the Guide to Authors the main body of the text. The Abstract form of journal clubs and people and should be written in complete sentences Generalviews. It also includes editorials on Policy without headings and should provide a

The format of the manuscript should be as which may include Intellectual property, summary not exceeding 250 words follows: Title page, Abstract, Introduction, Technology commercialization and inter- suitable for publication without the full M e t h o d s , R e s u l t s , D i s c u s s i o n , disciplinary issues. Journal is committed article text. Thus, if references must be Acknowledgements, Abbreviations, to the rapid publication of original cited in the Abstract they must include the References, Tables, Figure Legends and findings that increase our understanding author(s), journal title, volume number, Figures. Review articles are not required to of the molecular structure, genetics, page range, and year. It should begin by follow this outline. First mentions of function, behavior, physiology, toxicology discussing background, methods, results figures and tables in the text should be in and development of the nervous system. and conclusions but without these numerical order. Headings and sub-Experimental papers should have headers. The use of abbreviations in the headings should not end with a full stop. implications for the neurological disorders abstract should be avoided. Statistical The manuscript should be formatted in and may report results using any of a results need not be described in the double spacing and the lines should variety of approaches including anatomy, Abstract.not be numbered. The text of your theory, biophysics, invitro systems, Introductionpaper should be saved as a .doc file. imaging, and molecular biology. Papers

The body of the Research Articles should with the corresponding author’s last i n ve s t iga t ing the phys io log i ca l start with a brief Introduction, not name and sent to the editor by e-mail. mechanisms underlying pathologies of exceeding 450 words, which outlines the Manuscripts are expected to document the nervous system, or papers that report historical origins of the study and stating the origin and specificity of reagents used, novel technologies of interest to the aim of the study and/or hypothesis to particularly antibodies and document researchers in neurosciences are also be tested. It should not have information institutional authorization for conducting welcome. The criteria for acceptance of from Abstract or discuss the results.research in humans and animals, to manuscripts will be scientific excellence,

conduct adequate statistical analyses and originality, defiance of dogma based on Methodscomprehensively report statistical results, sound evidence and relevance to the field The methods section should be written and to be written in English.of neuroscience based on peer review of such that another researcher is able to

manuscripts. Although independent peer reproduce your work. Important Articles and Reviews could also be reviewers are requested by Editor, the methodological aspects of your work, accompanied with represen-tative colour authors may suggest a list of three names should be described, even if such photograph or illustrations which with expertise to comment on the paper descriptions can also be found in prior summarizes the work. This will be and upto two people who need to be publications. Companies from which displayed in the content.excluded from review. The editorial board materials were obtained should be listed

Title Pagereserves the right to refuse review process with their country.if the paper does not meet the scope of The Title page must include: Discussionthe journal or is not scientifically

? A clear and concise title The Discussion could contain the appealing. We encourage authors to summary of the major findings, while ? The authors name(s) and define the established principles and avoiding repetition of the statements in surnamespolicies that defy the norms and to argue Abstract or the Results. Citing literature against accepted dogma. The Editor in ? The address(es) from which the supporting or defying the findings should Chief will make a final decision on the work originated be provided. The Discussion should be acceptance of the manuscript and would able to project a major advancement of ? The name, address, fax number be under no obligation to seek further scientific concept, method or effect.and e-mail address of the person opinions. Our intent is that this

w h o w i l l d e a l w i t h Referencespublication becomes a gateway through correspondence, including which a wide array of information can be All references must be listed, and all listed proofs. accessed in a single periodical. The goal is references must be cited at least once in

to provide peer-reviewed and rapid ? The total number of pages, the main text.

Mission and Scope of Journal

Preparation of the manuscript Molecular Shots statement that the experiments were JPEG Images from experiments such as undertaken with the written consent of Research Articleimmunocyto chemistry can be submitted each subject, and that the study conforms

The research article should be typed in with legends and authorship. with institute ethical committee Calibri font size 12, double space word guidelines.Competing interestsformat, readable in Word 2003-07 format.

(ii) Studies involving experimental animalsIf there is likely to be economic gain The length of the article should not exceed (patents , f inanc ia l investments , 5000 words and all the illustrations should The methods section must briefly state consulting agreements) resulting from be in Power point edition 2007 or more. measures which were taken to minimize having utilized experimental approaches, The supplementary information, if any, pain or discomfort and the ethical instruments, methods or drugs, or of should be provided with Tables in word committee should be identified. having discussed such items in the format while all figures must be included

Experimental animalsmanuscript, represents a competing in power point. From a research article the references should be written as shown interest. Competing interests should be The species, strain, sex, age, supplier below. clearly disclosed and described in detail in and numbers of animals used should be

the Acknowledgements. It is in the specified. If transgenic or knockout From a Research Articleinterest of the author to disclose such animals are used the established

Jaime C. Montoya, Grace O., et al. interests, and ANS requires such nomenclatures must be usedDiagnosis of Tuberculous meningitis using disclosure.Enzyme-Linked Immunosorbent Assay Reagents

Fundingutilizing a 30,000-Dalton native antigen of The control experiments that were All sources of funding should be declared Mycobacterium tuberculosis. Phi J conducted to ensure the specificity of the in the Acknowledgements. I f a Microbiol Infect Dis 2000; 29 (4): 156-161. method should be described, along with private/commercial sponsor supported key references to previous work with this From a Chapter in a bookthe research, authors are advised to r e a g e n t . F o r a n t i b o d i e s , t h i s

e.g. Fullplus BW. Characterisation, describe the role of the study sponsor(s), if documentation includes a precise isolation and purification of cholinergic any, in study design; in the collection, description of the antigen, the nature of receptors. In : Theselff S Ed., Motor analysis, and interpretation of data; in the the antibody (species, purification), the innervation of muscle, 2nd ed. Longdon: writing of the report; and in the decision supplier, catalogue number, and Academic Press 1976; 11-26. to submit the paper for publication. If the specificity tests performed Review Articles funding source had no such involvement,

Statistical methodsthis should be stated.Review Articles in ANS are full length A complete description of statistical articles on topics of particular current Evaluation of manuscripts methods is required. The main statistical interest. Proposals for Review Articles are Submitted manuscripts are assigned to an results should be described in the Results invited by the Editors-in-Chief, Executive Associate Editor and/or and Editorial section. The description of statistical Editor and the Associate Editors. Journal Board member who is responsible for its results in the figure legends should be also encourages independent submission evaluation. He may get it evaluated from limited to important analysis. The of innovative and comprehensive reviews another expert in the field. The Editor-in- description of the statistical results should that provide complete framework of the Chief's decision regarding publication is include the degrees of freedom. topic including the historical aspects, based on the recommendation of the

Acknowledgementsprogress in the f ie ld inc luding reports of reviewers, which will, at the The financial support should be controversies and promises. The review Editors' discretion, be transmitted to the acknowledged alongwith those from should not simply provide an outline of authors.collaborators or if the author has worked every point published by a paper but

Ethical standards on a fellowship, the sponsoring agency rather be structured to address a point in should be mentioned. If the manuscript science covered the Journal. All studies using human or animal has been previewed by someone, the subjects should include carry a Case Reporti n d i v i d u a l ' s n a m e s h o u l d b e declaration identifying the Institution Only one case report of novel and extra acknowledged.or Review Committee which approved ordinary importance will be published.

the study. Editors reserve the right to Cover illustrationsUnstructured abstracts also required.reject papers if there is doubt whether Colour illustrations suitable for front

Journal Club appropriate procedures were followed cover, can be submitted to the Editor for Graduate students or post doctoral fellow or competing financial interests have consideration.can write a commentary or a published not been disclosed. Online submissionpaper from any reputed neuroscience

(I) Studies with human subjects ANS would be shortly launching the journal and describe the fundings under When human subjects are used, online submission of manuscripts at background study design, criticism and

implication. manuscripts must be accompanied by a the new web site.

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Official Journal of Indian Academy of Neurosciences

VOLUME l 16 APRIL 2009l l

AZAD HIND STORES

34/17-E, Chd. Telefax : 2704511-14Multicolour Offset Printers & Manufacturer of Computer Stationery

Cover Illustration of Neuronal & glial cell distribution in mouse hipppocampus (from Nagy Lab, LEFT), GFAP immunostaining of mouse cortical cultures (G. Joshi et al., TOP RIGHT) and Neurons stained with fluorescent cholera toxin B (from Preeti Lab, BOTTOM RIGHT)

advances and trendsEditorialNeel Kamal Sharma, Sudesh Prabhakar 45and Akshay Anand

Mini ReviewNew Focus72 Ovarian hormones and the brain

46 merit, seniority and sciencesignals

Suptendra Nath SarabadhikariB.W. Gawali, P.B. Rokade, G.B. Janvale and S.C. Mehrotra

Commentary48 asking the right question – the Report

ignored key of medical research75 3rd Canadian IBRO school of

Nusrat Shafiq neuroscience : neurodegeneration and regeneration

Research Article Akshay Anand49 A modified, rapid fluorometric

method for the quantification of Case Reportmitochondrial calcium

77 Dissemination of tubercular bacilli by Amit S. Korde and William F. Maragosshunt placement

Sandeep Mohindra, Rahul Gupta and Journal ClubRajesh Chhabra

54 Cells previously identified as retinal stem cells are pigmented ciliary epithelial cells Book ReviewPNAS 2009 ; 106 : 6685-6690 79 Animal Cell Technology Chandrika Abburi by Asok Mukhopadhyay

S.N. MukhopadhyayClassics

56 Mind – Body dichotomy Molecular ShotsVinod Srivastava Immunostaining pictures

57 Discourse on the method of rightly 80 Gururaj Joshi and Jeffrey A Johnson

conducting the reason and 81 Nibedita Lenka, Hoon-Ki Sung and seeking truth in the sciences

Paria Mohseni(Contd. from Previous Issue)

Rene Descartes

People and ViewsComprehensive Review 82 The new team

62 Age related macular degeneration - 95 Letters to Editor


Akshay Anand

Annals of Neurosciences

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