PHYSIOLOGYNEWS

winter 2006 | number 65

Meetings Bristol Brazil Heidelberg International Workshop, Kiev

Also featuring A century of research in sugar transport A summer in the life of a retiring physiologist Did evolution go the wrong path for the human lung? Acute pain Legs pay out for cost of breathing! Long and tortuous goodbye to Stud Muffin No 1

A publication of The Physiological Society IMAGES OF BRAZIL JOINT INTERNATIONAL MEETING WITH THE BRAZILIAN PHYSIOLOGICAL SOCIETY Ribeirão Preto, Brazil 27-30 August 2006

For more images of Brazil, including flora and fauna, see the inside back cover (photos by Prem Kumar) PHYSIOLOGYNEWS Editorial 3 Meetings Bristol Focused Meeting Johannes Reul, Astrid Linthorst, 4 The Society’s dog. ‘Rudolf Magnus gave Stafford Lightman me to Charles Sherrington, who gave me Heidelberg Focused Meeting Joseph Bruton 5 to Henry Dale, who gave me to The Images of Brazil inside front and back covers Physiological Society in October 1942’ Living history The surface energy of water from 1908 Wilfred Widdas 7 A summer in the life of ... Published quarterly by The Physiological Society An (endless?) summer in the life of a retiring physiologist 11 Contributions and Queries David Miller Executive Editor Features Linda Rimmer The Physiological Society Publications Office What structures bear the tension and store energy in lengthening 13 P O Box 502 muscle? Gavin Pinniger, K W Ranatunga, Gerald Offer Cambridge CB1 0AL Acidification protects skeletal muscle volume during anaerobic 15 UK exercise Juliet Usher-Smith, James Fraser, Christopher Huang Tel: +44 (0)1223 400180 Did evolution go the wrong path for the human lung? 17 Fax: +44 (0)1223 246858 John West Email: [email protected] Organization of intermediary metabolism – lactate and its 19 The Society web server: http://www.physoc.org transporters George Brooks, Takeshi Hashimoto Endothelial nitric oxide synthase-derived NO signals regulate 21 Magazine Editorial Board Editor microvascular permeability Walter Durán, Takuya Hatakeyama, Austin Elliott (University of Manchester) Fabiola Sánchez Members Acute pain Peggy Mason 23 Lesley Anson (Nature) Legs pay out for cost of breathing! Lee Romer, Jerome Dempsey 25 Patricia de Winter (King’s College London) Sarah Hall (Cardiff University) Functional domains of tropomyosin: new insights Robert Gaffin 27 Munir Hussain (University of Liverpool) Mariappan Muthuchamy John Lee (Rotherham General Hospital) International News Thelma Lovick (University of Birmingham) Bill Winlow (Prime Medica, Knutsford/University of Kiev International Workshop Bridget Lumb, Anne King, Oleg 29 Liverpool) Krishtal, Nana Voitenko, Zita Puskar, Volodymyr Khmyz, Oleksandr Maximyuk © 2006 The Physiological Society ISSN 1476-7996 The Society’s journals The Journal of Physiology Carol Huxley 35 The Society permits the single copying of Experimental Physiology Emma Ward 36 individual articles for private study or research. For copying or reproduction for any other purpose, Affiliate News written permission must be sought from The Young scientists and the media Selina Pearson 37 Physiological Society ([email protected]). From molecules to behaviour Izumi Fukunaga, Clare Howarth, 37 Opinions expressed in articles and letters submitted Rosemary Milton, Ede Rancz by, or commissioned from, Members, Affiliates or Memorable Members outside bodies are not necessarily those of The David Henry Smyth Ian Roddie 38 Physiological Society. The Physiological Society Lectures The Physiological Society is registered in England The Bayliss-Starling Prize Lecture Tim Biscoe 39 as a company limited by guarantee: No 323575. Registered office: PO Box 11319, London WC1X Book Review 41 8WQ Parliamentary and Scientific Committee 42 Registered Charity: No 211585. Carol Huxley Printed by The Lavenham Press Ltd Parliamentary Links Day Liz Bell 43 Cover photos Society News Governance for beginners – the structure of The Society 45 Simon Kellas Society rebranding Donna Brown 46 Scientific Meetings Calendar 2007 47 Lister Institute Research Prizes 48 Biosciences Federation Richard Dyer 49 Images of Brazil Unbelievable! (front cover by Nana Voitenko back cover by Prem Kumar) Long and tortuous goodbye to Stud Muffin No 1 50 Obituaries Douglas Ingram Joy Dauncey 51 PHYSIOLOGYNEWS

Action points Guidelines for contributors In this issue Grants These guidelines are intended to assist Welcome to our final Physiology News For full information on Members’ authors in writing their contributions and to of 2006. and Affiliates’ Travel Grants, reduce the subsequent editing process. The Network Interaction Grants, Non- Editorial Board of Physiology News tries to A lot of biology has to do with energy. Society Symposia Grants, Vacation ensure that all articles are written in a The word ‘energy’ sometimes gets journalistic style so that they will have an Studentship Scheme, Departmental appropriated by pseudo-science and immediate interest value for a wide Seminar Scheme, Centres of readership and will be readable and used for the purposes of mystification, Excellence and Junior Fellowships comprehensible to non-experts. In but this should not detract from the visit: http://www.physoc.org/grants particular, scientific articles should give a central role of energy in science. In good overview of a field rather than focus biological systems the conversion of Membership applications entirely on the authors’ own research. energy from one form to another – e.g. Applications for Full and Affiliate chemical energy from ATP to muscle Membership are received throughout Format of articles The main message or question posed should be work, or to ion pumping – is a central the year and have no deadlines. A introduced in the first paragraph. The background part of what physiologists have been decision is normally made within 7 for the topic should then be established, leading investigating over the last 200 years. days of the Administration Office up to the final conclusion. receiving the application. For full This issue contains a fair bit of energy. Length of articles details please visit: This will be determined by the subject matter and Wilfred Widdas makes a plea for more http://www.physoc.org/join agreed with the Executive Editor. attention for surface energy as a phenomenon in Living History, while Submission of articles Change of address Authors should submit articles as a Word muscle, the energy-transducing Members should inform the document attached to an email. Illustrations machine that has fascinated Administration Office of any changes should be sent as separate attachments (see below) physiologists perhaps more than any of address, telephone, fax or email and not embedded in the text. other, gets a good look, with four News address. Illustrations and authors’ photographs and Views pieces and a meeting report Changes can be emailed to: Authors are encouraged to submit diagrams, from Heidelberg. [email protected] or updated online drawings, photographs or other artwork with their articles or to suggest appropriate illustrations. A At this time of year the decreasing daily at http://www.physoc.org photograph of the author(s) should also accompany submissions, if possible. Illustrations hours of solar energy inevitably tend to and photographs may be colour or black and be a source of some regret. If you feel white, prints, transparencies or tif/jpeg files with a this way, perhaps the pictures from the minimum resolution of 300 dpi. Electronic colour figures should be saved in CMYK mode. recent meeting in tropical Brazil may Physiology News cheer you up. And finally, even the sun Deadlines References setting (on a career) can have its Authors are requested to keep the number of upsides, as David Miller explains in his Letters and articles and all other references to a minimum – preferably no more contributions for inclusion in the than two or three. Please cite all references in the diary of a retiring physiologist. Spring 2007 issue, No. 66, should style of The Journal of Physiology (see reach the Publications Office Instructions to Authors 2005 at Austin Elliott ([email protected]) by 21 http://www.physoc.org) December 2006. Short news items are encouraged, and can usually be included as late copy if space permits.

Suggestions for articles Suggestions for future articles are welcome. Please contact either the Executive Editor or a member of the Editorial Board of Physiology News (see contents page for details).

Physiology News Online Physiology News is now available on The Society’s web site: http://www.physoc.org. See Unbelievable! (p. 50) EDITORIAL PN 3

The homeopaths, and the companies that acupuncture, or the physiological effects of Homeopathic ‘mumbo­ produce over-the-counter homeopathic alterations in diet. remedies, are understandably delighted. jumbo’ But scientists want evidence, not anecdotes Well, you might say, so what? The placebo and hand-waving. If proper science shows real Many scientists these days have at least the effect is not new, and a fool and his/her money physiological effects, beyond those of a odd moment when they feel that their view of are soon parted. Most scientists would agree placebo compound or sham intervention, and if the world is under threat from a tide of what the that the labelling is a joke, but in a world awash these can be made to work as a treatment, journalist Francis Wheen, in his best-selling with ridiculous claims, why get worked up? what you have is a therapy. Rather than being book, termed ‘mumbo-jumbo’ (1). a question of ‘alternative’ or ‘mainstream’, it is Well, firstly, perhaps, because the MHRA, down to what works – or more precisely, what For scientists, ‘mumbo-jumbo’ manifests itself acting on our behalf, is supposed to care – we can be sure works because it can be shown in the rejection of scientific understanding of their web site states they ‘enhance and to work in a properly-designed scientific how the physical world works in favour of safeguard the health of the public by ensuring experiment. mystical beliefs derived from a range of that medicines and medical devices work, and sources. are acceptably safe.’ (7). How they reconcile Which highlights something else we should be the first part of this statement with the change thinking about – our failure, as professional The examples are too numerous to list, but as in the homeopathy rules is not clear. scientists, to inform enough of the public about the debate swirls it occasionally coalesces what proper controls are, and exactly why around particular issues. Recent flash-points some experiments are convincing, and others are the challenge to evolutionary theory from ‘The Physiological Society is concerned with are not. About what the placebo effect is in so-called ‘Intelligent Design’ (2), and the the scientific investigation of how the body medical experiments and trials. About what question of whether complementary and works … It is our view that "alternative homeopathy actually is – you would be alternative medicine has any scientific basis. medicine" has, with very few exceptions, no surprised how many people, including a good scientific foundation, either empirical or few bioscience graduates I have met, think it Homeopathy has recently taken centre stage in theoretical. As an extreme example, many means ‘herbal remedies’ rather than ‘infinitely this latter debate. In May an open letter signed homeopathic medicines contain no dilute nothing’ – and why it is scientifically by 12 senior doctors and scientists (including molecules of their ingredient, so they can nonsensical. several Physiological Society Members) urged have no effect (beyond that of a placebo). that alternative therapies unsupported by To claim otherwise it would be necessary to None of these is terribly complex to explain, evidence of efficacy should not be used in the abandon the entire molecular basis of and many of them go to the heart of what NHS (3). Later, the demonstration that chemistry. The Society believes that any science is, and how it is done. In many ways, homeopathic pharmacies advised homeopathic claim made for a medicine must be based this seems a golden opportunity to use the formulations, which have no antimalarial action, on evidence, and that it is a duty of the public interest to put across how science offers as malaria prophylaxis garnered national media regulatory authorities to ensure that this is a clear way to divide what actually works from attention (4). done.’ what doesn’t.

Following the homeopathic malaria remedy As the immortal Richard Feynman put it: media exposé, it might be thought that the Secondly, because – at the risk of sounding ‘Science is a way of trying not to fool yourself. scientific absurdity of homeopathy had been incredibly pompous – there is a principle at The first principle is that you must not fool clearly demonstrated. Imagine, therefore, the stake, namely that decisions of this kind should yourself, and you are the easiest person to surprise of many scientists and doctors when be made on the basis of scientific and medical fool.’ the Medicines and Healthcare Products evidence and understanding. Regulatory Agency (MHRA), the body charged As this issue went to press, The Society’s with controlling the safety of medicines in the Finally, the MHRA’s decision to allow licencing statement, along with many others, was being UK, decided to allow homeopathic remedies to and sale of homeopathic remedies in this way passed to interested members of the House of be sold with packaging featuring – for the first is likely to be widely interpreted as approval of Lords in advance of a debate on the new time in 30 years – claims about what the alternative remedies in general. This in turn will homeopathy regulations on 26 October. By the remedy purports to treat (5). foster the perception that they work. time you read this, we should know if it did any good. But whatever the result, get polishing Previously a homeopathic remedy in a high The Physiological Society, like other scientific your homeopathy-debunking speech. And I like street chemist would have been labelled ‘6c societies, has been asked by the lobby group to think Feynman would not mind us pinching dilution of Gelsemium sempervirens’, or Sense About Science (5) to comment on the his lines. something similarly obscure. It can now be MHRA decision and has issued a statement sold, quite legally, as ‘NoCold-Max cold and flu reaffirming its belief in scientific evidence, and Austin Elliott remedy … homeopathic’. decisions based on it.

1 Wheen F (2004). How mumbo-jumbo conquered the world: a short history And as the web site of the European Council The Society’s statement (box) is not, note, a of modern delusions. Harper Perennial. for Classical Homeopathy (6) puts it: ‘To make blanket dismissal of all the things the public 2 http://www.royalsoc.ac.uk/news.asp?year=&id=4298 such a claim, the manufacturers need only commonly regard as complementary therapies. 3 http://www.timesonline.co.uk/article/0,,8122-2191985,00.html show that the product has been used to treat Physiologists have long studied the effects of 4 http://news.bbc.co.uk/2/hi/uk_news/5178488.stm those particular conditions within the exercise upon the body, and the physiological 5 http://www.senseaboutscience.org.uk/index.php/site/project/86 homeopathic industry.’ No scientific basis. No actions of plant-derived substances. Work goes 6 http://www.homeopathy-ecch.org/ clinical trials. No evidence of effectiveness. on into the possible physiological basis of 7 http://www.mhra.gov.uk/home/idcplg?IdcService=SS_GET_PAGE&nodeId=5

Physiology News | No. 65 | Winter 2006 | www.physoc.org 4 PN EDITORIAL

Physiology News | No. 59 | Summer 2005 | www.physoc.org 4 PN BRISTOL FOCUSED MEETING

New developments in stress physiology – from gene to man A Physiological Society Focused Meeting at the University of Bristol on 4 and 5 December 2006

interactions, variants of the glucocorticoid receptor and interactions between the glucocorticoid receptor and MAPK and NFkB. This will be Stafford Lightman (above), followed by a discussion of the Astrid Linthorst (right, top) regulation of the HPA axis at and Johannes Reul (right). hypothalamic and higher brain levels and how targeted mutations of The Meeting on New developments in glucocorticoid receptors can alter stress physiology – from gene to man feedback and, in turn, the stress will provide a broad view of stress and response. The next session will look at glucocorticoid physiology from the the effects of stress on neuronal most basic cell and molecular biology plasticity and include work on cell through neuronal plasticity to adhesion molecules, dendritic and integrative regulation and synaptic remodelling and perinatal glucocorticoid action in disease. programming. Finally, there will be a session on glucocorticoid actions in The first session of this Physiological health and disease which will cover Society Focused Meeting will discuss stress, obesity, pregnancy and aspects of glucocorticoid – chromatin psychiatric disease.

Invited speakers In addition to the main sessions there will be oral communications from Trevor K Archer submitted abstracts whilst the other Julia Buckingham abstracts will be presented as posters. Mary F Dallman E Ronald de Kloet The Meeting will also host the Bayliss- Alison Douglas Starling Prize Lecture* which will be Gordon Hager presented by Rod Flower and James P Herman introduced by Society President Ole Isabella Heuser Petersen. Colin Ingram Krisztina J Kovacs The conference dinner will be aboard Louis J Muglia the historic s.s. Great Britain and we Michael Norman are all looking forward to and Pier Vincenzo Piazza expecting a really exciting and From the top: A view of the University with the Wills David Ray convivial meeting. Memorial Building Tower in the foreground; The Johannes M H M Reul Dorothy Hodgkin Building, Focused Meeting venue; Benno Roozendaal SS Great Britain, venue for the Meeting dinner; Johannes Reul, Astrid Window artwork in the Dorothy Hodgkin Building. Carmen Sandi Linthorst and Stafford Jonathan R Seckl Lightman *See Tim Biscoe’s history of the Bayliss- Michael G Stewart Organisers, University of Bristol, UK Starling Prize Lecture on p. 39.

Physiology News | No. 65 | Winter 2006 | www.physoc.org HEIDELBERG FOCUSED MEETING PN 5

Control and modification of excitation-contraction coupling in healthy and diseased muscle

The Medical Biophysics Unit of the Ruprecht-Karls University of Heidelberg hosted this Physiological Society Focused Meeting on 13 September 2006. Nick Boross-Toby from The Society ensured the smooth organization together with Oliver Friedrich, Rainer Fink and Sally Davison for the hosts. The location was the splendid Alte Aula – a long, narrow high-ceilinged heavily wooded hall. Its north-south orientation with the bright sun beating on the windows meant that the late afternoon would have been very conducive to sleep were it not for some exciting selected oral presentations in the last session.

Hans Christoph Lüttgau, who had recently celebrated his 80th birthday was a guest of honour and homage was paid to him by many of the symposium speakers. It was pleasing too to have Caspar Rüegg and Reinhard Rüdel with us at the symposium.

The morning session, with Martin Schneider, Werner Melzer and Eduardo Ríos, focused on fundamental release events in skeletal muscle and elements of their control. The bottom line was that, despite huge leaps over the past few years, we still lack vital detail. After a coffee break and our first encounter with the posters in a bright and airy room, it was then back into the solemn hall. Godfrey Smith showed us that after coronary artery ligation, epicardial conduction thought the scar tissue with its surprisingly rich perfusion is essentially unchanged

Left (from top) Time to admire the Alte Aula at the end of the day. Charalambos Sigalas after his oral communication. Sun beating down on the entrance to Alte Aula. Right (from top) Ole Nielsen with Juliet Usher-Smith, who was awarded the best poster prize. Vincent Jacquemond (left), Peter Szentesi and Janos Almassy at lunch. Hans Christoph Lûttgau, doyen of skeletal muscle physiologists. George Stephenson at the poster session. James Fraser and Gerald Elliott at lunch.

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Speakers Martin Schneider (University of Maryland, USA) Werner Melzer (University of Ulm, Germany) Eduardo Ríos (Rush University, Chicago, USA) Godfrey Smith (University of Glasgow, UK) Manuela Zaccolo (Venetian Institute for Molecular Medicine, Italy) Angela Dulhunty (Australian National University) George Stephenson (La Trobe University, Australia) indicating that groups of myocytes presumably survive in the infarct area. Manuela Zaccolo persuasively presented evidence for the role of phosphodiesterase 2, 3 and 4 isoforms in the compartmentalization of cAMP/PKA signalling.

A surprising tasty light lunch recharged glycogen stores. The first speaker in the afternoon, Angela Dulhunty, described the consequences of the splice variants of the RYR1 and RYR2 receptors and mutations in calsequestrin on RYR opening and cellular Ca2+ homeostasis. George Stephenson was the final symposium speaker, and reported his recent work on the determinants of transverse tubule excitability in the mechanically skinned skeletal muscle fibre.

The final session was devoted to selected oral communications. Charalambos Sigalas told us of the complexities of calmodulin modulation of the RYR2 receptor and then Stephanie Reis reported on the subtle effects of R145G substitution in cardiac troponin I on rat cardiomyocyte contractility. Myotonia and the potential + mechanisms of [K ]o in electrical stabilization was addressed by Sunisa In between these two talks, James From the top Chaiklieng. Ole Nielsen used the Fraser presented a modelling study of Inside the Alte Aula, waiting for the final session of the mechanically skinned rat EDL fibre to day to start. the determinants of sarcoplasmic + + estimate the number of Na -K pumps reticulum electrochemical equilibria Martin Schneider giving his talk (left); Eduardo Ríos + answering questions after his presentation (right). and maximal clearance of K in the which aroused a surprisingly passionate transverse tubule. The same preparation Manuela Zaccolo at the start of her talk (left); Claude response from the audience. Collet (left) and Bruno Allard during the coffee/poster was used by Bradley Launikonis to session. 2+ show how store-operated Ca Joseph D Bruton George Stephenson (left), Bradley Launikonis and homeostasis is initiated and terminated. Karolinska Institute, Stockholm, Sweden Eduardo Ríos at the coffee break.

Physiology News | No. 65 | Winter 2006 | www.physoc.org LIVING HISTORY PN 7

The surface energy of water from 1908 Wilfred Widdas looks back on a century of research which set him on an unfashionable path in sugar transport, and emphasises the debt we owe to physiologists

I envisaged that the surface energy of Outlining my scheme and ideas at a water provided the power for the coffee morning break, interest was working of the intra-membranous shown by all those present, and my protein GLUT1. This protein was the colleague and former PhD student, red cell member of proteins that are Graham Baker (Fig. 1; Physiology now termed MFS proteins (major News 55, 49-50), made the most facultative superfamily).These positive suggestion when he said, ‘If membrane proteins enable water surface energy is doing the work of soluble molecules to cross lipid transport, then we should be able to membranes. The amino acid sequence slow down glucose exits by reducing of this membrane protein was described the surface tension, and we could by eight investigators in 1985 Figure 1. Wilfred Widdas (right) with the late Graham probably do that by using alcohols (Mueckler et al.). Their structure had Baker explaining their posters at the IUPS meeting in since red cells can withstand high 12 alpha helices, which they proposed Glasgow, 1993. concentrations of methanol’. spanned the lipid membrane. Several groups had proposed that the protein cellular systems is difficult to The glucose exits referred to had been a formed clefts which opened appreciate, but my main interest has form of study of human erythrocytes in alternatively to the outside to allow been the application of the surface a double beam photo-electric apparatus glucose to enter and then closed behind energy of water derived in evaporation/ with a 25 ml cuvette that had a stirrer it but opened a similar water-filled cleft condensation cycles at submicroscopic and 2 cm light path. Red cells to the inside, which allowed the sugar dimensions. incubated to hold 78-100 mM glucose to enter from its central position in the when injected into glucose -free saline membrane. This scheme would fit in Jumping forward nearly 50 years when made their presence felt by an initial with the carrier kinetics I advanced 50 working with a former PhD student at osmotic swelling, but this is rapidly years earlier when first studying sugar the merged Bedford and Royal followed by a linear osmotic shrinkage permeability at St Mary’s Hospital Holloway colleges, a crude model of as the glucose is lost. That the exit is Medical School under Professor the 12 alpha helices as the membrane linear for over 80% of the record on a Huggett. part of the GLUT1 protein convinced pen recorder was shown by earlier me that the scheme of working of the users of the apparatus. Traces in a Philosophically one can argue that life glucose transporter would only be publication by a former PhD student on this planet is supported by radiation possible if it was powered by the are shown in Fig. 2. Following from the sun. Short wave-length energy surface energy of water, which was free Graham’s suggestion, alcohols were is used for photosynthesis to produce of metabolism but different from found to equilibrate very fast and oxidisable chemicals from CO2 and Gibb’s free energy of proteins like ATP. records of glucose exits were found to water. Physical chemists know that water evaporates at all temperatures but that the longer wave-length energy replaces the latent heat energy lost from the oceans by the latent heat of evaporation of water. This radiation from the sun keeps the global energy in balance as a steady state. This is important for molecules that leave the water surface and enter the atmosphere as a gas and diffuse upward being lighter than oxygen and nitrogen in the air. The water vapour condensing in the clouds may fall as rain. The rain that falls does mechanical work in eroding rocks and soil, and transfers the salts and solutes to the rivers and seas. How the same cycles of evaporation/ condensation of water can be harnessed Figure 2. Linear exits of glucose from red cells with 78 mM glucose into saline’s with low glucose concentrations to do effective mechanical work in (from Sen .& Widdas, 1962).

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be just as linear. An example with ethanol at 7.5% is shown in Fig. 3. The inhibitory effect of alcohols was readily measured.

Soon after this we started on the experiments that Graham had suggested and found that the other lower alcohols reduced both the surface tension and inhibited glucose exit rates at even smaller concentrations than methanol. In the course of these studies, the four alcohols inhibited the glucose exits to 50% in concentrations that progressively reduced (Fig. 4). This almost threefold reduction in concen­ tration was in accord with a physical chemistry phenomenon, which was Figure 3. A tracing of a glucose exit from 100 mM glucose in 7 ½ % ethanol (2.28 M), which is four fold isotonic discovered by Traube in 1891 and is concentration but is rapidly equilibrated to give a linear 67% inhibited exit (from Widdas & Baker, 1991). known as Traube’s Rule (Findlay, 1944)

The linear fall in glucose exit rate with the percentage fall in surface tension from 100% to 65% of that of water, and the fit of the 50% inhibition values to Traube’s rule, were both pointers to the involvement of the surface energy of water as suggested from the knowledge of physical chemistry (Fig. 5). Further, we found that the proposal for the involvement of surface tension of water in biological functions was not original. Figure 4. A copy of the Table 2 taken from the same 1991 reference. This table records the concentration for 50 Czapek, a botanist from Jena had % inhibition of glucose exit ( by lowering of the surface tension) and the approximation to Traube’s Rule. A recent reported in 1911 that he had found that but second application of these alcohols to biological studies (Ly & Longo, 2004) has also given reductions in plants were not viable if the surface surface energy according to Traube’s Rule. tension was reduced to 68% that of normal water. This toxic value of lowered surface tension (68%) corresponded closely to the cut off value (65%) for glucose exits. The experimental results were reported to The Physiological Society in two pre­ circulated communications at the 1990 Bristol Meeting. One was given by Graham Baker and I presented the other. Both were approved, but a longer paper was rejected by two journals and only published over a year later by a less well known journal whose Advisory Editor was our Professor of Physical Chemistry (Widdas & Baker, 1991).

However, the structural model of the opening and closing cleft mechanism involved the breaking of the water channel to the liquid water at the centre, firstly from the outside (after the Figure 5. The linear fall of glucose exit rates as a percentage of the maximal rate (100%) plotted against the sugar had diffused into the centre from surface tension of the saline relative to normal water (100%). the outside bulk water). The separation

Physiology News | No. 65 | Winter 2006 | www.physoc.org LIVING HISTORY PN 9 allowed an outer bolus of water to be water surface eliminated multiplied by Thus, not only has the surface energy expelled and the cleft at this side the value of the energy content per unit of water not been accepted as closed. As spherical glucose sugar was area. For water this latter value is explaining the working of the glucose now in the centre, the 12 alpha helices already well known as 73 x 10-3 Jm-2. transporter protein in erythrocytes for were regarded as being semi-rigid and In the membrane transporter for the last 17 years, but its behaving like planks on a child’s glucose, the breaking of the continuous supplementation of the ATP-hydrolysis seesaw. They closed around the glucose water channel needs energy to create in muscle contraction, suggested by like they would around a fulcrum, and two half spheres for the two new ends Widdas & Baker in 2001 and again in this opened the cleft on the inside as a of water. Two halves are a whole, so 2004, is also ignored and unacceptable near vacuum. knowing the radius, the total area is to the established scientific opinions. 4πr2; and the energy is easily put into The opening as a near vacuum is due to heat quantities, or joules. When this The modern interest in genetics has two physical chemical principles, which was done it was found that having many examples where we find clefts few understand. The first is that the measured the turnover rate too small to admit liquid water but mouth of the inside cleft is far too radioactively, and also the number of involving detailed reactions that need a small to admit liquid water, which transporters in each red blood cell, it watery base. Their source of energy to would need Laplace’s pressure (P = 2 was possible to determine the total do detailed work outlined by earlier λ /r) and this would be over 2,000 mechanical work being done by the microscopists is never explained, but is atmospheres for a radius in the nm glucose transporters in 5 litres of blood presumed to follow from the structural dimensions. Consequently the water every minute. The value was more than detail published by Watson and Crick forms a plane surface across the mouth 10 times that of the resting metabolism (1953). Novel proteins are synthesised of the conical cleft. This is also seen in for an adult of 70 kg. from changes in DNA as part of umbrellas where the fabric is full of complicated chemical changes that small holes but the penetration of water This unexpected and surprising result clearly need more than simple thermal does not occur because of surface of physical chemical calculations had vibrations but without the work tension. The second principle is that a several philosophical implications. involved being fully explained. plane surface of water exposed either to Firstly, it was impossible to imagine air or to a vacuum space is a liquid that this amount of work free of If in future investigations physical surface from which individual water oxidative metabolism was provided chemists take more interest in molecules can leave by evaporation. only for the membrane transport of biological functions, studies of the The water vapour enters the cleft where glucose. During the long epochs of surface energy of water will feature it can condense on polar groups or on evolution nature would have found more strongly in the next decades of water molecules already present. more widespread uses for the work the 21st century. potential of the surface energy of water. The change from liquid to gas is a Work production is largest in muscles, Wilfred F Widdas change of state needing 540 calories which are naturally considered for a Honorary Member, Lincolnshire, UK per gram. It is difficult to visualise the system where the surface energy of enormous amount of extra energy water might be used. This had already References which is hidden away as latent (hidden) been proposed by Bernstein (1908) Bernstein J (1908). Uber die Temperaturkoeffizienten der Muskelenergie. Pflugers Arch 122, 129-195. energy in liquid water. The physical almost 100 years ago. chemists sometimes refer to this as Findlay A (1944). Introduction to Physical Chemistry. Longmans, mechanical energy since it is the only However, the provision of heat energy Green & Co, London. part of the surface energy of water that is not enough to explain mechanical Ly Hung V & Longo Marjorie L (2004). The influence of short-chain can do mechanical work (Widdas, work and therefore a description of the alcohols on interfacial tension, mechanical properties, area/molecule 1993). The First Law of muscle ‘engine’ was looked for in the and permeability of fluid lipid bilayers. Biophys J 87, 1013-1033. Thermodynamics explains how all literature, but found to be lacking. The Mueckler M, Caruso C, Baldwin SA, Panico M, Blench I, Morris HR, forms of energy have a heat equivalent, reason for the failure, is seen as the Allard WJ, Lienhard GE & Lodish HF (1985). Sequence and structure of a human glucose transporter. Science 229, 941-945. but the reverse is not true. A source of attempts to explain the muscle heat energy is not an explanation of contraction mechanism entirely from Sen AK & Widdas WF (1962). Determination of the temperature and pH dependence of glucose transfer across the human erythrocyte how work is done. To perform work an one energy source, namely that of ATP membrane measured by glucose exit. J Physiol 160, 392-403. ‘engine’ or ‘engine-equivalent’ is hydrolysis whereas the surface energy Watson JD & Crick HC (1953). Genetic implications of the structure required to convert the energy source of water was readily available as a of deoxyribosenucleic acid. Nature 171, 964-967. into meaningful or purposeful second energy source to supplement the Widdas WF & Baker GF (1991). The role of the surface energy of movements, or into some other form of ATP energy source. To provide a water in the conformational changes in the human erythrocyte work. scheme employing two energy sources glucose transporter. Cytobios 66, 179-204. in place of only one is much easier and Widdas WF (1993). Hidden work and energy. The Physiological In the case of work from surface a paper proposing this was submitted Society Magazine 11, 5-6. energy of water it is found that if you to Experimental Physiology but was Widdas WF & Baker GF (2001). The surface energy of water: the have an ‘engine’ the magnitude of work rejected as being unsuitable at this time largest but forgotten source of energy in biological systems. possible is proportional to the area of given its particular focus. Cytobios 106, 7-54.

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Wilfred Widdas recalls: maternal sheep only had glucose in their Special offer for blood, whereas the sheep fetus (the Physiological Society My entry to the London physiology scene unborn lamb) had glucose and fructose in Members came just after World War Two, 9 years its blood, so that its blood sugar was after I had qualified as a doctor. Following The motoneurone and its higher than its mother’s. I was able to muscle fibres 1 year in general practice, I had spent show from the results of Huggett and his seven and a half years in the Royal Army biochemist, Warren, that the glucose could The Physiological Society Medical Corps, being released with the not cross the placenta by simple diffusion Monograph No 50 by Daniel Kernell honorary rank of Major. Seeking a post­ (as stated in the most prestigious – War job in science, I was rejected by the American – biochemistry textbook of the Secretary of the MRC and the Head of the day, but must come across by a special National Institute for Medical Research at ‘carrier’ mechanism. Mill Hill. Professor Jo Bernal, the Government adviser in physical research, The mathematics I used were beyond could only offer me a sociology job, living Huggett’s mathematical expertise, so he in a new town and reporting on how the sent my calculations to A V Hill for advice. East Enders adjusted to living in their new As I remember, Hill replied that it was environment – at least, that was how I outside his field but he had been advised interpreted it. It was quite opposite to my by his biochemist colleague, E J Harris, ideas of scientific research work, though, that diffusion could be the explanation and so I declined. therefore recommended rejection of my approach. I was too junior to engage Hill After some other unsuitable options, I was any further and did not obtain Harris’ lucky to get one of five postgraduate explanation as to how diffusion could scholarships offered by St Mary’s Hospital explain the concentrations I had used in Medical School for ex-service doctors and my analysis. started to work under Professor A St G Huggett, later FRS. These scholarships Although I cannot remember the exact Buy now from Oxford University were worth £500 per year, much less than dates (these Living History articles often my army salary had been but the grant Press and take advantage of an start ‘I clearly remember’ – for myself this exclusive 25% discount was free of tax. I accepted the effective would be impossible. My memory is good 10 years ‘juniority’ relative to the other for some items but for others it has gone, This book, published in August normal research students for 3 years and time scale in particular is often far 2006, presents a state-of-the-art before being given a lectureship. from clear. Perhaps from the perspective summary of knowledge concerning of 90+ years precise dates are not so the motoneurones, summarising both Huggett had come from Leeds as important!), this must have delayed me by classical and recent knowledge. This professor, and brought with him his chief about 2 years until I found that R B Fisher is the first time that so many aspects technician Mr Hancock – in my opinion the and Parsons, biochemists at Oxford, were of this broad subject have been best physiological technician in London at using the same formulae as I had used for treated in one comprehensive the time. A V Hill’s chief technician, Mr glucose transport in other situations. So I monograph. Parkinson, who was the expert at making presented two papers in the early 1950s. the special thermocouples for A V’s Hardback, 360 pages. research, was also very good and was the When I had collected data from my own Physiological Society Members only one of Hill’s Department whom I knew blood as well as that of fetal cells, I wrote Price £41.25 (RRP £55.00), plus well. Parkinson and I shared collecting a PhD thesis. Huggett got Professor F J postage and packing. Government surplus apparatus for W Roughton of Cambridge to act as research after the cessation of hostilities. external examiner. At the interview, I recall To order, please telephone We had to visit empty RAF hangars where that Roughtson said, ‘Widdas, I have read +44 (0) 1536 741727, or email equipment would be displayed for sale to through your thesis and recalculated all [email protected]. Please ‘junk shops’ in Tottenham Court Road. the equations in it. I found them all quote the code AMPHYSOC06 Luckily, universities had a special ‘first correct. But … (he paused, and you can when ordering to obtain the chance’ to buy. I got an ink-writing pen imagine my anxiety rising), then he discount. recording milliammeter, which I used for continued ‘but, I found three sentences many years. without a verb in them.’ Professor Offer ends 31 March 2007. Roughton, FRS, had hit on a weakness I At St Mary’s the research was centred on have admitted since school days and now Offer prices are only valid when fetal physiology – copied from Huggett by I usually get a relative with a BA in English ordering direct from Oxford Sir Joseph Barcroft at Cambridge as a Literature, or similar, to read my papers University Press and cannot be wider study. The interesting fact that set (presently this is my grand-daughter who redeemed in bookshops. me on a path in sugar transport was that has a BA from Lancaster University).

Physiology News | No. 65 | Winter 2006 | www.physoc.org A SUMMER IN THE LIFE OF ... PN 11

An (endless?) summer in the life of a retiring physiologist David Miller enjoys the nostalgia and rewards of an early retirement from university life

As with my predecessors in this series, this is weeks/months/summers rather than A day in the life of… To set the scene, this spring I took the too­ tempting early retirement deal from Glasgow University; my transitional state is my introspective preoccupation.

In late April came the standard retirement ‘do’. (I’d started at Glasgow on 1 April 1977 but – to paraphrase The Who – wouldn’t get fooled again by leaving on the 1 April too). In my case, the ‘do’ included a valedictory seminar and a late afternoon gathering of colleagues, with several surprise visitors, then speeches and many ‘farewells’ gratefully received. Fellow retirees will recognise the growing Figure 1 (above). Hunter’s Moon in Torshavn harbour, June 2006. sense I had of being at my own funeral Figure 2 (below(). Neil Spurway and David Miller moored at Midvagur, Faroes. as the unjustified words of praise and thanks were heaped up. ‘inherited’ from my late colleague some active and prominent in Oliver Holmes. Tilly Tansey, at the physiology, some in medicine (i.e. The rites of passage to retirement have Wellcome Trust, told me that even the applied physiology), others hardly ever continued since then with clearing my complete J Physiol she assembled for heard from, or of, since. I recalled Reg office and downsizing the lab. This the digitising project could not be Chapman’s remark to me on process (ongoing) generated dozens of found a home, so I despair of ‘saving’ completion of my own thesis that ‘he bags of confidential waste, general my modest sub-set. Debating the merits had now replaced himself, so his job waste, recycling waste and a long of print browsing versus the was done’. (However, like many ‘help-yourself’ bookshelf for oxymoronic, electronic ‘internet supervisors, Reg didn’t stop after his bibliophile colleagues to plunder. browsing’ is for another place. first scientific ‘child’. Just what is a However, I still wrestle with one reasonable number for one’s scientific widespread problem; how to deal with My clear-out also fuelled a powerful progeny?) 80 shelf–feet of The Journal of surge of nostalgia. I was sharply Physiology? I have a run from 1961 to reminded of many past colleagues, of Stimulated by reading some of the old date, the volumes prior to 1978 students (undergrad and postgrad), exam papers, lecture notes, and committee papers that choked office and lab, I was driven to ponder the changes wrought on our science and university education. If you have any to hand, do glance at some 1970s, 80s and 90s past exam papers. They will surely reveal the ‘dumbing down’ that 30 years (or more) have wrought. Notwithstanding the broader curriculum that many courses have deliberately adopted, the intellectual pitch has declined. The best students do remain excellent but the range of student attainment is much wider now so median standards are surely lower. Yet, paradoxically, median ‘grades’ are higher. Ponder the standard of Honours research projects. While (mere) technical sophistication rises steadily, the aspiration to novelty and

Physiology News | No. 65 | Winter 2006 | www.physoc.org 12 PN A SUMMER IN THE LIFE OF ... thoroughness seems much lower. was available in June to crew for my Research unveiled at The Assuming the experiments work, the long-time Glasgow colleague Neil Physiological Society’s Main ease with which data can be collected, Spurway, sailing to the Faroes in his Meeting held at University analysed and presented now is stunning 31-foot boat Hunter’s Moon (Figure 1 College London from 4 to 7 compared with what earlier decades and 2). I joined Neil in Orkney. July 2006 allowed. Yet the conversion of data into Fortuitous timing saw a day as an results and the quality of the final External Examiner for the Aberdeen Obesity risk for children can be traced to a mother’s diet output seems not to match the new medics end for me at the harbour, rather New research, funded by the EU’s early tools available. Do you detect a whiff than the railway station, as I boarded nutrition programming project, reveals that a of the disenchantment that contributed the ferry to Kirkwall. Over the next mother’s intake of a high fat diet during to my early retirement decision? week, Neil and I sailed, with sight­ pregnancy could lead to persistent changes seeing interruptions, through Orkney to in a child’s bodyweight leading to obesity in Amongst special trophies that my clear­ Fair Isle (which lived up to its name) adulthood. The research, carried out by out has generated, the nostalgia wave and thence to Shetland. A few days Anne-Maj Samuelsson (Division of rose high as I chanced upon delights there set us up for the near–200 mile Reproduction and Endocrinology, King’s such as: log and trig tables, the passage from Ronas Voe (NW College London), investigated the ‘Rubber’ Handbook, my Castell Shetland) to Torshavn (the Faroes’ mechanisms whereby a mother’s diet ‘engineering’ slide rule, three cycle log capital). We completed the run in about influences the offspring. The research graph paper, 5.25” floppy discs, a 34 hours, almost entirely on one tack reveals that a mother’s intake of a fat-rich Sinclair programmable calculator thanks to a remarkably steady, if at diet during pregnancy is more likely to owner’s manual, Letraset, and much times uncomfortably strong, south­ produce offspring with increased fat stores more. I have now ceremonially handed westerly wind, emboldened by the due to enlarged fat cells, altered fat over my cherished box of fine Allan reassuring ‘advice’ of GPS. I stayed a metabolism, and higher lipid level in their keys to Godfrey Smith’s safe hands. He further week aboard as we explored blood. is one of a dwindling group who knows much of what this unique Atlantic what they are and has good uses for archipelago has to offer (except The tight-fit brain – study reveals them. But, despite a ruthless clear-out, Guillemot, which despite the 600,000 brain swelling as a cause of migraine headaches the ‘bottom drawer’ is still full of nesting pairs available, was The secrets of migraine headaches may lie results to be written up, biographical unaccountably ‘off’ one restaurant’s in acute mountain sickness where brain notes on Sydney Ringer and more, plus menu that week). So what do you know swelling as a result of lack of oxygen I have the stimulation of my of the Faroese Nobel Prize winner for causes severe headache and vegetative membership of The Society’s History Physiology Niels R Finsen? (For me, symptoms. The challenge associated with and Archives Committee to help to fill the return journey was a simple 1 hour understanding headaches is that academics my days from here on. And my wife, flight. Neil S made it safely back and usually study the symptoms once they have Carolyn, has ‘a list’. still afloat, if not entirely without formed. Research led by Damian Bailey incident, some days later). (University of Glamorgan) highlighted for Amongst the responsibilities that I have the first time what happens during the passed on, one concerns the advent of My ‘real’ summer holiday was to NE evolution of a headache. Volunteers were ‘electronic’ PhD theses. My last 5 years Spain and SW France, and it did exposed to lack of oxygen in a simulated involved postgraduate administrative prompt one physiological train of high altitude chamber, to investigate the matters as head of the Faculty Graduate thought. Might the acclimatisation to build up to, and wind down from, the School. This led me to be volunteered higher temperatures on the Med, the resulting headaches. Under these as the sole academic on the steering attendant dehydration and my raised conditions (at a simulated altitude of group of EThOs (Electronic Theses daily alcohol intake be associated? A 4,600m), half of the subjects typically Online), a national project concerned significant proportion of alcohol developed symptoms that were with making theses searchable and dehydrogenase activity is inducible and indistinguishable from a migraine. accessible on line.* I raised there the alcohol is a diuretic: could the concerns about shared intellectual acceleration of C2H5OH breakdown as Heat halts pain inside the body property, plagiarism, self-censorship a holiday proceeds be a major Brian King (UCL) led research that found and priority that, if neglected, will dog contribution to reduced diuresis and the molecular basis for the long-standing this otherwise laudable aspiration. At thus ‘rehydration’? I reckon after 2 theory that heat, such as that from a hot­ root, a profound reappraisal of the very weeks I’m usually just about water bottle applied to the skin, provides nature of the UK PhD thesis, never equilibrated. If anyone has acted as an relief from internal pains, such as stomach mind the training and education of the alcohol-free control for this aches, for up to an hour. The team found PhD itself, seems long overdue. phenomenon, do let me know. that the heat receptor, known as TRPV1, can block P2X3 pain receptors, thus On an entirely happy note, my personal David Miller stopping the pain being sensed by the body. transition from ‘work’ to ‘play’ was Honorary Research Fellow, Biomedical & Life marked by a wonderful opportunity. I Sciences, University of Glasgow, UK More information at http://www.physoc.org *http://www.gla.ac.uk/ibls/NBS/nbsstaff/djmiller/etheses.rtf (News and Events)

Physiology News | No. 65 | Winter 2006 | www.physoc.org FEATURES PN 13

What structures bear the tension and store energy in lengthening muscle? When active muscles are stretched the cross-bridge cycle is truncated by suppressing the power stroke and thereby reducing ATP consumption; the high tension they exert is generated by stretch-induced strain in the pre-stroke cross-bridges. Other, non­ crossbridge, structures also make a substantial contribution to tension and energy storage

Textbooks of physiology often give the actin (step 2). Tension is generated by a impression that when muscles are power stroke, a conformational change activated to produce tension, they either in the myosin head, associated with shorten performing work or remain at release of Pi (step 3). Detachment of constant length if restrained by a post-stroke heads, after they have lost suitable load or antagonistic muscle ADP and rebound ATP, completes the (isometric contraction). Yet, stretch of cycle (step 4). How does this cross­ an active muscle, somewhat bridge cycle adapt to lengthening? confusingly referred to as an eccentric Since the power stroke generates force, contraction, occurs frequently during it is strain-sensitive and hence it is daily activities. Examples include the expected to be slowed by the increased lengthening of the biceps when Gerald Offer (left), KW Ranatunga and Gavin Pinniger. strain on the heads caused by the lowering a heavy weight, or of the calf lengthening. muscles when walking downstairs. isometric tension. Yet the consumption Lengthening contractions may also of the fuel, ATP, and hence the energy So a simple explanation for the serve as a braking mechanism to output (heat plus work) declines. What reduction of ATP consumption in decelerate the body when running is the molecular explanation for this lengthening muscle is that the cross­ downhill or landing from a jump. seeming paradox? It will be recalled bridge cycle is truncated, the power­ Lengthening muscles also store energy that the structural basis of muscle stroke and subsequent detachment of which may be used during a subsequent contraction is the cross-bridge cycle, post-stroke heads being increasingly shortening contraction to enhance the the interaction of heads of the myosin shut down as the lengthening velocity tension produced; this phenomenon is molecules (M) of the thick filaments is increased. But if the power stroke the underlying basis of plyometric with actin (A) filaments. The Lymn- rate is reduced, how do we account for exercise-training. Yet our understanding Taylor scheme summarises the main the increased tension exhibited during of the molecular processes involved in events in this cycle (Fig. 1A). lengthening? The pre-stroke heads that lengthening has been unclear. Hydrolysis of ATP occurs on myosin attach to actin, given a sufficient heads detached from actin (step 1). lifetime, will be dragged to higher An active muscle strongly resists being The heads carrying the products of strains and this will produce force; the stretched, developing up to twice the hydrolysis, ADP and Pi, then attach to heads will eventually detach when highly strained and reattach to actin subunits further away from the Z-disc (Lombardi & Piazzesi, 1990; Fig. 1B). It had been supposed that a special mechanism (forcible detachment) might operate when tension prised the pre­ stroke heads off the actin but it is now appreciated that when force is applied to a complex of two proteins, it merely enhances the spontaneous rate of dissociation. So tension can be generated during lengthening without the power stroke, the high force being borne by pre-stroke, rather than post­ stroke heads (Getz et al. 1998; Pinniger et al. 2006). Our computer simulations Figure 1. Lymn-Taylor cross-bridge cycle and its truncation during lengthening A, In isometric muscle: Step 1 is ATP hydrolysis on the detached heads, step 2 is the attachment of myosin of active muscle based on the Lymn- heads to actin or their detachment; step 3 is the power stroke (or its reversal) involving the swing of the lever arm Taylor scheme are able to account for of the myosin head; and step 4 is the irreversible step. Note that in isometric muscle the filament positions remain the lengthening limb, as well as the the same and force is generated by the power stroke which stretches the elastic element, here schematically shortening limb, of the force-velocity represented by a spring connecting the myosin head with the thick filament backbone. B, During lengthening: The pre-stroke heads initially attach to the actin subunit marked with an asterix. The relation without the need to postulate movement of the actin filament from left to right stretches the spring increasing the force and the heads detach at extra intermediates in the cross-bridge high strain. They reattach at an actin subunit further from the Z-disc and the process is repeated. cycle.

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So far we have discussed only the Figure 2. Tension responses contribution that cross-bridges make to to stretch and contribution of cross-bridges and non­ tension. But there are strong crossbridges to tension indications that other, non-crossbridge, A, Tension response (upper structures also contribute (Edman & trace) from a rat fibre bundle Tsuchiya, 1996). For example, during to a 5% ramp stretch (lower lengthening, when work is done on the trace) applied on the plateau of an isometric tetanic muscle, much of the energy absorption contraction. Also shown are cannot be explained simply by the two isometric contractions, enhanced cross-bridge strain (Linari et one at the initial length and al. 2003). Two further indications are the other at the extended length, and the tension shown by the tension responses during response in resting muscle. and after the application of a constant B, Tension response at an velocity stretch to a bundle of expanded time scale to show isometrically contracting muscle fibres the change in slope on the (Fig. 2). There is an initial rapid phase tension rise (denoted by of tension rise due to the cross-bridges arrow). being increasingly strained, interrupted C, The effect of velocity on the peak tension, the early on by a small but detectable dip contribution to the peak in tension as the post-stroke heads tension by cross-bridges and undergo the reverse power stroke. the residual tension. The There is then a marked reduction in the non-crossbridge tension is the difference between the rate of tension rise (Fig 2B); peak tension and the simulations suggest that this transition crossbridge tension. (indicated by the arrow in Fig. 2B) is due to the cross-bridges approaching (BTS), the rapid phase of tension rise of Ca2+ on activation and play a the new steady-state truncated cycle during stretch and the fast phase of significant role in energy storage. (Fig. 1B). However, the tension tension decay afterwards are greatly continues to rise slowly thereafter to diminished, whereas the slow phases of Acknowledgements the end of the stretch. After the stretch, tension rise and decay and the residual We thank the Wellcome Trust for tension decays to a level which is force remain high (Pinniger et al. support and Hamish Roots for help higher than the isometric tension. 2006). Our interpretation is that non­ with illustrations. cross-bridge structural elements stiffen Both this continuing slow tension rise on activation and provide a tension that Gavin Pinniger1,2 after the transition and this residual increases progressively throughout the K W Ranatunga1 force enhancement are hard to explain stretch. The peak tension thus has Gerald Offer1 on the basis of cross-bridges acting contributions from both cross-bridge 1 alone and have been attributed to a Muscle Contraction Group, Department of Physiology, and non-crossbridge elements; the School of Medical Sciences, University of Bristol, non-cross-bridge component. The peak residual force enhancement arises from Bristol, BS8 1TD, UK tension increases with velocity of a partially decayed non-crossbridge 2current address is School of Biomedical and Chemical stretch to a plateau of about twice the force. Sciences, The University of Western Australia, isometric force, whereas the residual Crawley, WA 6009, Australia. force enhancement remains the same The residual force enhancement and irrespective of velocity. Figure 2C continued slow tension rise during a References illustrates the velocity dependence of stretch had been attributed to sarcomere Edman KA & Tsuchiya T (1996). Strain of passive elements during force enhancement by stretch in frog muscle fibres. J Physiol 490, the contribution to tension by cross­ inhomogeneity and sarcomere popping, 191-205. bridges (measured by the tension at the uncontrolled elongation of sarcomeres. Getz EB, Cooke R & Lehman SL (1998). Phase transition in force transition) and the contribution by non­ However, no convincing direct during ramp stretches of skeletal muscle. Biophys J 75, 2971-2983. cross-bridge elements. The cross­ evidence of sarcomere popping has Linari M, Woledge RC & Curtin NA (2003). Energy storage during bridge tension rises steeply to a plateau been observed experimentally and, stretch of active single fibres from frog skeletal muscle. J Physiol 60% higher than the isometric force at tellingly, no popping is observed when 548, 461-474. high velocity, whereas the non-cross­ active myofibrils are stretched even Lombardi V & Piazzesi G (1990). The contractile response during bridge tension is insensitive to velocity. though their tension response to stretch steady lengthening of stimulated frog muscle fibres. J Physiol 431, Two further findings support the shows the above features (Telley et al. 141-171. existence of these two tension 2006). Our proposition is that Pinniger GJ, Ranatunga KW & Offer GW (2006). Crossbridge and components. Firstly, the tension decay interactions between actin and the titin non-crossbridge contributions to tension in lengthening muscle: force-induced reversal of the power stroke. J Physiol 573, 627-643. after ramp stretch also shows fast and filaments that connect thick filaments slow phases. Secondly, when active to the Z-disc, or between actin and C­ Telley A, Stehle R, Ranatunga KW, Pfitzer G, Stüssi E & Denoth J (2006). Dynamic behaviour of half-sarcomeres during and after muscle force is drastically reduced by a protein (an accessory protein of thick stretch in activated psoas myofibrils: sarcomere asymmetry but no specific myosin-crossbridge inhibitor filaments) are recruited by the release “sarcomere popping” J Physiol 573, 173-185.

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Acidification protects skeletal muscle volume during anaerobic exercise Biophysical measurements and theoretical modelling of resting skeletal muscle exposed to extracellular lactate demonstrate that intracellular buffering of hydrogen ions precisely offsets the effect of increased intracellular lactate on cell volume

Muscle exercise has long been A associated with the transmembrane entry of water and an accompanying cell swelling (Kjellmer, 1964; Lannergren, 1990). Although it is now generally accepted that this water movement is the result of an increase in intracellular osmolarity (Ward et al. 1996), the precise origin and nature of this increase in osmolarity is not well understood. One candidate often implicated is the intracellular lactate produced during anaerobic metabolism. However, lactate accumulation is then B accompanied by production of H+ ions from the hydrolysis of ATP and recent charge-difference modelling of skeletal muscle electrophysiology (Fraser & Huang, 2004), which suggested that steady state cell volume is determined principally by the intracellular membrane-impermeant anion content and its mean charge valency, predicted that cellular acidification would produce quantifiable reductions in cell volume through the association of H+ ions with these impermeant anions. This hypothesis was subsequently confirmed when experiments producing intracellular acidification in amphibian muscle through addition and subsequent withdrawal of NH4Cl from the extracellular solution (Fraser et al. 2005) produced predictable decreases in cell volume that were proportionate to the increase in intracellular [H+]. This suggests a mechanism by which cell volume could be conserved in the face of the production of lactate and H+ during anaerobic exercise. Thus, whilst the additional lactate would increase the intracellular osmolarity and so tend to increase cell volume, the accompanying acidification would be expected to reduce cell volume. The consequent steady-state volume would then depend on the balance between Figure 1. A, Plot of the mean cell volume change, ∆Vc, (± S.E.M, n = 6) against time during and following these two opposing effects. exposure to an isosmotic solution containing 80 mM lactate. B, Charge-difference modelling of the addition at 15 min and withdrawal at 45 min of isosmotic 80 mM lactate Ringer solution from a model cell initially at steady state Our study (Usher-Smith et al. 2006) in normal Ringer solution. Horizontal bars at 45 min show the final steady-state values of each modelled variable after more extended exposures to lactate-containing solutions, while horizontal bars from 100 min similarly denote establishes that, in fast-twitch final values after extended periods of recovery in normal Ringer solution. Thus charge-difference modelling amphibian muscle, these two effects predicted that lactate addition and withdrawal would cause minimal volume change, despite an increase of - precisely balance each other. By [lactate ]i to >15 mM during the lactate loading phase, in accordance with the experimental results depicted in A.

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experimental study but nevertheless important to the mechanisms underlying these adjustments. For Figure 2. Charge-difference example, it was possible to examine the modelling of volume changes effect of variations in total intracellular over time during exposure to buffering capacity on the volume isosmotic 80 mM lactate Ringer for cells with intracellular changes (Fig. 2). This showed that, as buffering capacities of (from top expected, the small initial volume down) 0.5, 0.9, 1.1 and 1.5 increase observed on exposure to times normal. Horizontal bars extracellular lactate is not affected but plotted past 50 min show final the final steady state volume is stable steady state values of Vc in each case. profoundly influenced by the relative intracellular buffering capacity: exposing resting frog muscle to lived and, whilst still exposed to addition of lactate and H+ to a cell in extracellular solutions containing solutions containing lactate, the cell which pHi has no influence on cell lactate we were able to reproduce the volume returned to its control value. volume due to a relative buffering + increases in intracellular lactate and H Isolated increases in intracellular lactate capacity of zero produces a 13% + observed following exercise whilst and H to levels comparable with those volume increase whilst 80 mM avoiding the wide range of other observed following stimulation do not, extracellular lactate would be expected metabolic and osmotic alterations therefore, significantly alter steady-state to produce a 7% volume decrease if which occur during exercise. An cell volume. intracellular buffering capacity was integration of the results from a range increased by a factor of 1.5. It is only A modified charge difference model of of experimental and theoretical with buffering capacities similar to the quantitative relationship between techniques then enabled us to those actually measured in frog muscle volume and membrane potential quantitatively examine the combined that the final volume is not significantly developed by Fraser and Huang (2004) effects of these increases in lactate and different to control values as seen + then combined these individual cellular H on cell volume. experimentally. observations to reconstruct the osmotic and electrophysiological workings of By using 1H-NMR to compare the Acknowledgements the cellular system as a whole. In intracellular lactate concentration, We thank our collaborators Julian Griffin agreement with the experimental and Peter Bailey (both at the University of [lactate] , resulting from exposure to i findings, simulated addition of Cambridge). This work was supported by extracellular lactate at a range of extracellular lactate produced the Medical Research Council, the concentrations with that following low­ intracellular lactate accumulation, Wellcome Trust, the Royal Society and frequency electrical fatiguing cellular acidification and a transient Astra Zeneca with additional support from stimulation, we successfully developed the James Baird and George Henry Lewes volume increase followed by recovery a protocol that increased [lactate] in Funds. i of the volume to initial resting values resting muscles to levels close to those (Fig. 1B). It also clarified the following fatigue. Corresponding mechanism responsible for these Juliet A Usher-Smith measurements of intracellular pH, pH , i volume changes. The initial increase in James A Fraser using pH-sensitive microelectrodes cell volume could be attributed to a Christopher L-H Huang similarly confirmed that the pHi in rapid entry of lactate and resultant Department of Physiology, Development and Neuroscience, University of Cambridge, UK these resting muscles was within the increase in intracellular osmolarity but, range previously recorded in fatigued as the pH subsequently decreases and References amphibian muscle. Relative cell volume + H ions bind to negatively charged sites Fraser JA & Huang CL-H (2004). A quantitative analysis of cell was then measured using confocal xz­ on intracellular membrane-impermeant volume and resting potential determination and regulation in plane scanning during and after excitable cells. J Physiol 559, 459-478. anions, the charge on those anions, z , exposure of resting fibres to lactate x becomes less negative. This results in a Fraser JA, Middlebrook CE, Usher-Smith JA, Schwiening CJ & (Fig. 1A). Addition of extracellular Huang CL-H (2005). The effect of intracellular acidification on the decrease in the net intracellular anionic lactate produced an initial increase in relationship between cell volume and membrane potential in charge and so allows potassium, the amphibian skeletal muscle. J Physiol 563, 745-764. cell volume but this swelling was short- predominant intracellular cation, to Kjellmer I (1964). The effect of exercise on the vascular bed of leave the cell, reducing the intracellular skeletal muscle. Acta Physiol Scand 62, 18-30. osmolarity and returning the cell Lannergren J (1990). Volume changes of isolated Xenopus muscle volume towards control values. fibres associated with repeated tetanic contractions. J Physiol 420, 116P.

This close agreement between Usher-Smith JA, Fraser JA, Bailey PSJ, Griffin JL & Huang CL-H (2006). The influence of intracellular lactate and H+ on cell volume in experimental and theoretical results amphibian skeletal muscle. J Physiol 573, 799-818. additionally prompted an extension of the model to allow analysis of cellular Ward DS, Hamilton MT & Watson PD (1996). Measurement of tissue Juliet Usher-Smith (left), James Fraser and volume during non-steady state high-intensity muscle contraction. Christopher Huang. parameters not directly amenable to Am J Physiol 271, R1682-1690.

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Did evolution go the wrong path for the human lung? Birds and mammals evolved along two separate evolutionary lines from the primitive reptiles. Although the physiology of many of the organ systems of birds and mammals are similar, the lungs are radically different. The thesis here is that the bird ended up with a better lung

play no part in gas exchange. The gas is into the lung, the gas cannot reach the then moved through the gas-exchanging peripheral alveoli by convection, and parabronchi where the pulmonary the last part of its movement is by a capillaries are located, and gas complicated combination of convection exchange occurs. In other words, this is and diffusion in the small airways. As a a flow-through system rather like the result, the more proximal alveoli tend radiator of a car, and in fact gas to be better ventilated than the more exchange is very similar in many distal, resulting in stratified inequality respects to heat exchange. Furthermore, of ventilation. This inequality is small although it is not obvious from Fig. 1, at rest but increases on exercise when the gas flow through the parabronchi is the time available for diffusion is less. John West unidirectional as a result of The flow-through system in the bird aerodynamic valving in the airways, largely avoids this problem. Although it is customary to emphasize although the details of this are still not the ways in which the structure and fully understood. 2. The pool pattern of ventilation function of the human lung are well­ reduces the alveolar PO2 suited to its mission of gas exchange, The reciprocating nature of ventilation Because a small amount of inspired gas the opposite case can be made. The in mammals results in at least three is delivered to a large pool of alveolar proposition here is that the avian lung potential problems. gas, the PO2 of the latter is substantially is superior in many respects to the below the inspired value. For example, mammalian lung. 1. Potential for uneven ventilation in the human lung, the inspired PO2 is In resting man, the inspired tidal about 150 mm Hg but the PO2 of the It was about 300 million years ago that volume of about 0.5 litres is delivered alveolar gas and arterial blood is the ancestors of the present reptiles into a lung volume of about 3 litres. around 100 mm Hg. Again the flow­ emerged from water and made a Although the geometry of the airways through system of the bird largely commitment to air breathing, but they allows the inspired gas to penetrate far avoids this problem too. were exothermic and could not sustain high levels of exercise. However, from them came the two great evolutionary lines that produced the mammals and birds, both capable of high, sustained levels of oxygen consumption. A fascinating aspect of these two evolutionary paths is that the physiology of the nervous, cardiovascular, renal, gastro-intestinal, endocrine and skeletal muscle systems show many similarities, but the lungs are radically different. The contention here is that the bird lung is superior to that of the mammal and that evolution got off on the wrong track.

Figure 1 shows the anatomy of the avian and mammalian lungs. Ventilation in the latter is reciprocating with the relatively small tidal volume of each breath being delivered to a relatively large volume of gas already in the lung. In the bird the arrangement is quite different. Inspired air is drawn into air sacs which are avascular and Figure 1 Diagram of the bird and mammalian lungs that both evolved from ancestors of the present reptiles.

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Figure 2 Typical microscopical sections of bird lung (A) and mammalian lungs (B). In the bird the pulmonary capillaries are supported by a dense honeycomb-like network of air capillaries. In the mammalian lung the pulmonary capillaries are spread out along the free-floating alveolar wall.

3. The reciprocating pattern of the blood-gas barrier and thus current pattern of gas exchange, which necessitates large terminal air units interferes with diffusive gas exchange. tends to increase the PO2 of blood Because, as already stated, gas leaving the lung. This is well movement to the most peripheral The mammalian lung has other recognized and has been fully discussed alveoli is by a combination of disadvantages compared with the avian elsewhere (Piiper & Scheid, 1972). convection and diffusion, the terminal lung but these can only be touched on air spaces need to be relatively large to briefly here. As Fig. 1 indicates, the Finally, as mammals, we tend to regard keep the resistance low. The striking bird has successfully separated the gas­ ourselves as at the top of the differences between the gas-exchanging exchange function of the lung from its evolutionary heap, but this is arguable. tissue of the avian and mammalian ventilatory function. The Some birds are superior to any lungs are shown in Figure 2. The blood bioengineering requirements of the mammals in their mass specific oxygen capillaries are similar in size but the air tissues for these two functions are very consumption and their aerobic scope, spaces are dramatically different. The different. Gas exchange by diffusion and there are more species of birds bird has air capillaries that branch off necessitates an extremely thin blood­ (about 9,000) than mammals (about the parabronchi or their extensions and gas barrier, for example, it is only 0.2­ 4,200). their diameter is about 10 to 20 µm 0.3 µm thick over much of the area of So why did evolution proceed along an depending on the species. By contrast the barrier in the human lung. But the apparently flawed path for mammals. the alveoli of the human lung, for repetitive movement required by This is probably not a useful question. example, have a diameter of about ventilation is better done by the more Evolution does not have a goal but 300 µm. robust air sacs. In fact, one of the commonest, serious lung diseases in proceeds incrementally. From time to Some of the disadvantages of these humans is emphysema which is time there is a change in few base pairs relatively large alveoli have only characterized by breakdown of the which confer an advantage or recently been appreciated. Figure 2B delicate alveolar walls. disadvantage for survival. But the fact shows that the capillaries are strung out that birds have better lungs than along the alveolar wall rather like a Another very practical disadvantage of ourselves helps to keep a sense of string of beads. This means that they the mammalian lung is that occlusion perspective. have no mechanical support at right of an airway often causes serious Acknowledgements angles to the alveolar wall. As a result impairment of gas exchange. This is a The experimental studies were made the capillary wall has to be thicker. In very common problem in the with Zhenxing Fu and Rebecca Watson. particular, the extracellular matrix of postoperative setting where airways are The work was supported by NIH grant the blood-gas barrier, which is largely blocked by retained secretions or RO1 HL 60968. responsible for it strength, needs to be aspiration of fluid. Because the same very much thicker than in the bird structures are responsible for both John B West where the pulmonary capillaries are ventilation and gas exchange, this is a Department of Medicine, University of California San supported by a network of surrounding much more serious problem for Diego, La Jolla, CA, USA air capillaries (West et al. 2006). mammals than is presumably the case Furthermore, the mammalian alveolar in birds, where aspirated material goes References walls require a cable of type 1 collagen into the nonvascular air sacs. Piiper J & Scheid P (1972). Maximum gas transfer efficacy of models for fish gills, avian lungs and mammalian lungs. Respir Physiol 14, that threads its way between the 115-124. capillaries to maintain the integrity of Another advantage of the avian lung is West JB, Watson RR & Fu Z (2006). The honeycomb-like structure the relatively large and unsupported that the arrangement of air and blood of the bird lung allows a uniquely thin blood-gas barrier. Respir alveolar wall. This again thickens part capillaries results in a so-called cross­ Physiol Neurobiol 152, 115-118.

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Organization of intermediary metabolism – lactate and its transporters

Once thought to be a ‘dead-end’ metabolite produced because of O2 insufficiency, lactate is produced continuously under fully aerobic conditions in humans and other mammals. This production serves several functions, key of which is that it is permissive of glycolysis. Lactate is removed mainly by oxidation in red skeletal and cardiac muscle fibres, but also by the liver and kidneys where lactate is the major gluconeogenic precursor. By affecting redox in cells as well as in cell compartments of Takeshi Hashimoto (left) and George Brooks with a poster outside the laboratory removal, lactate serves as a signalling molecule. Lactate traverses cell that lactate is the consequence of O2­ significant is that lessons learned on membranes by means of several limited metabolism in humans or other muscle can be generalized to the monocarboxylate (lactate/pyruvate) mammals.’ Rather, the O2 debt ideas functioning of other cells and tissues, transport proteins (MCT). Our recent are traceable to early 20th century including brain (Brooks, 1998). work using microscopy and a variety of studies on non-circulated and non­ techniques shows that MCTs and oxygenated amphibian muscle Based on isotope tracer, arterial-venous related proteins occupy cell domains to preparations. But now, based on the difference mass balance and biopsy facilitate the exchange and use of recent report of Hashimoto et al. studies (Brooks et al. 1991) we now lactate. (2005), as well as related prior know that skeletal muscle is not only investigations, it is time to turn over the the major site of lactate production, but Lactate is an end product of classical idea of lactate production due also the major site of its removal, glycogenolysis and glycolysis produced to O2 lack. Today, we can think of mainly via oxidation. Lactate and only when cells lack oxygen: ‘this I lactate as a substrate for mitochondrial pyruvate are exchanged across muscle learned during my undergraduate respiration that can be shared among cell (sarcolemmal) membranes by studies in Japan’, said Takeshi cell compartments, tissues and organs. facilitated, proton-linked transport Hashimoto. ‘Well, that’s what I learned As well, lactate serves as a (Roth & Brooks, 1990) involving a also as an undergraduate in the 1960s’, gluconeogenic precursor, and as a family of MCT proteins (Garcia et al. replied George Brooks. ‘But’, Brooks signalling molecule, a ‘lactormone’ 1994). MCT1 is widely expressed in went on to add, ‘as a graduate student because of its influence on redox. And, different tissues and has been localized and ever since, I could not find proof perhaps physiologically more in muscle to sarcolemmal and mitochondrial membranes (Brooks et al. 1999; Hashimoto et al. 2005), and facilitates uptake of lactate from interstitium and plasma. The putative role of MCT4 is cellular lactate extrusion. Up to now, Brooks and associates have posited two major lactate shuttles:

• the cell-to-cell lactate shuttle: lactate formed in some muscle cells with high rates of glycolysis can shuttle to other cells with high oxidative capacity via sarcolemmal lactate transporters and be oxidized (Brooks, 1998)

Figure 1. Serial cryostat sections stained for MCT1 (A) and MCT4 (B). Fibre type identifications were made on • the intracellular lactate shuttle (ILS): the bases of myosin ATPase, SDH, and α-GPD staining (SO, thick arrows; FOG, thin arrows; FG). Very little lactate enters mitochondria and is MCT1 is visible in FG fibres, either at sarcolemmal or interfibrillar cell domains. MCT4 was concentrated at the oxidized to pyruvate via mitochondrial cell surface in fast (FOG and FG) fibres. Note that in SO fibres MCT1 is strongly stained, while little MCT4 is detected. Scale bar = 50 µm. The quantitative analysis of MCT1 (a) and MCT4 (b) protein expression in terms of lactate dehydrogenase (LDH) when optical density (OD) in different fibre types are shown. Values are expressed means ± SE. **P<0.01. mitochondrial redox decreases, and

Physiology News | No. 65 | Winter 2006 | www.physoc.org 20 PN FEATURES pyruvate is oxidized via the tricarboxylic acid cycle and electron transport chain (Brooks et al. 1999).

However, the existence of mitochondrial MCT1 and LDH has been controversial. Until our recent paper (Hashimoto et al. 2005), no one had quantified MCT isoform expression by histochemical assessment. Consequently, it was unclear whether MCTs occupied the same or different cell domains in different muscle fibre types. To better understand the physiological roles of MCTs and related proteins, such as mitochondrial inner membrane protein cytochrome oxidase (COX), we sought to determine the distribution and relative abundances of MCTs in rat plantaris that contains different muscle fibre types. Quantitative immunohistochemical Figure 2. Cellular locations of A, MCT1, CD147 and COX, and B, COX and LDH determined using confocal laser determination of MCTs by the high scanning microscopy (CLSM) and fluorescent probes for the respective proteins. In A, MCT1 was detected sensitivity avidin-biotin complex throughout the cells including subsarcolemmal and interfibrillar domains of oxidative fibres (arrows: plate a and c). (ABC) method showed that MCT1 is CD147, anchoring protein of MCT1 (plate b) is localized corresponding to MCT1 (plate a). When these MCT1 located at the sarcolemma and (green), CD147 (red), and COX (blue) were merged, superposition of the two probes was clear (white: plate d). In B, when these COX (green) and LDH (red) were merged, superposition of the two probes was clear (yellow: plate throughout the cell interior in oxidative c). Scale bar = 20 µm. fibres; in contrast, MCT4 is highly expressed in the sarcolemmal domain localizing it to the cell surface. Like Brooks’ sons Dan and Tim (now 29 and of glycolytic fibres (Fig. 1). As well, MCT1 (Fig. 2A-a), we detected CD147 27 years old) grew up with O2 debt confocal laser-scanning microscopy throughout rat plantaris muscle fibres theory in the biology curriculum. But, (CLSM) demonstrated that MCT1 and (Fig. 2A-b). Furthermore, MCT1 (Fig. will Kengo Hashimoto (18 months old) COX are co-localized at both 2A-a), CD147 (Fig. 2A-b), and COX have the same experience? interfibrillar and subsarcolemmal cell (Fig. 2A-c) were co-localized in domains (Fig. 2A). These results show individual fibres of rat plantaris muscle George A Brooks that MCTs and associated proteins are (Fig. 2A-d). These findings indicate Takeshi Hashimoto positioned to facilitate function of the the existence of MCT1 at mitochondrial Integrative Biology lactate shuttles: lactate formed in some inner membrane associated with University of California, Berkeley, CA, USA muscle cells with high rates of CD147. Furthermore, we showed the References glycolysis (e.g. fast-glycolytic, FG co-localization of COX (Fig. 2B-a) and Brooks GA (1998). Mammalian fuel utilization during sustained fibres) could be readily released via LDH (Fig. 2B-b) as a yellow colour in exercise. Comp Biochem Physiol B Biochem Mol Biol 120, 89-107. MCT4 (and MCT1) and transported Fig. 2B-c, indicating the existence of Brooks GA, Butterfield GE, Wolfe RR, Groves BM, Mazzeo RS, into slow and fast oxidative (SO and LDH in mitochondria. Additionally, in Sutton JR, Wolfel EE & Reeves JT (1991). Decreased reliance on FOG) fibres via sarcolemmal MCT1 cultured L6 skeletal muscle cells, we lactate during exercise after acclimatization to 4,300 m. J Appl Physiol 71, 333-341. lactate transporters. In conjunction, also observed associations among mitochondrial MCT1 facilitates lactate mitochondrial MCT1, CD147, COX Brooks GA, Dubouchaud H, Brown M, Sicurello JP & Butz CE (1999). Role of mitochondrial lactate dehydrogenase and lactate uptake, as oxidation is the major means and LDH by both CLSM and oxidation in the intracellular lactate shuttle. Proc Natl Acad Sci U S A of intramuscular lactate disposal and immunoprecipitation technique 96, 1129-1134. therefore essential in establishing the (Hashimoto et al. 2006). Although not Garcia CK, Goldstein JL, Pathak RK, Anderson RG & Brown MS concentration and pH gradients that yet definitive, results are consistent (1994). Molecular characterization of a membrane transporter for lactate, pyruvate, and other monocarboxylates: implications for the drive cellular lactate release, uptake and with presence of a sixth, lactate Cori cycle. Cell 76, 865-873. disposal. oxidation, mitochondrial complex. Hashimoto T, Masuda S, Taguchi S & Brooks GA (2005). Immunohistochemical Analysis Of Mct1, Mct2 And Mct4 Expression As to the controversy surrounding So, what is the pace of progress? In Rat Plantaris Muscle. J Physiol 567, 121-129. mitochondrial MCT1 and LDH, we Takeshi Hashimoto learned classic ‘O 2 Hashimoto T, Hussien R & Brooks GA (2006). Colocalization of provide strong evidence of their debt’ theory in the late 1990s. Now, MCT1, CD147 and LDH in mitochondrial inner membrane of L6 cells: existence. The single-span checking PubMed there are hundreds of evidence of a mitochondrial lactate oxidation complet. Am J Physiol Endocrinol Metab 290 1237-1244. transmembrane glycoprotein CD147 hits for ‘MCT1’ and the ‘lactate (BSG, or Basigin) is considered to be shuttle’, where much recent activity has Roth DA & Brooks GA (1990). Lactate transport is mediated by a membrane-bound carrier in rat skeletal muscle sarcolemmal the chaperone protein for MCT1 been in the realm of neurobiology. vesicles. Arch Biochem Biophys 279, 377-385.

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Endothelial nitric oxide synthase-derived NO signals regulate microvascular permeability Regulation of vascular tone is largely dependent on the production of nitric oxide (NO) by the endothelium. Recent findings provide compelling evidence supporting an important role for endothelial NO in the regulation of microvascular permeability

necessarily compelling as it was based intensity (IOI) using fluorescently mainly on the ability of L-arginine labelled dextran (FITC-dextran 70, with analogs to block NOS non-specifically. mean molecular weight = 70,000 daltons) as a macromolecular indicator. The development of genetically Topical application of PAF caused an engineered mice has contributed increment from a baseline value of importantly to evaluate more directly 2.4 ± 2.2 to a peak net value of the function of eNOS in microvascular 84.4 ± 2.7 units in wild-type mice; Walter Durán (above, permeability, with the appropriate whereas in mice lacking the eNOS gene left), Takuya Hatakeyama (above, caveats related to the adaptive changes (eNOS-/- mice) PAF increased IOI only right) and occurring in response to deletion of from a baseline of 1.0 ± 0.3 to a peak Fabiola Sánchez (left) important genes. Using eNOS-/- mice, net value of 15.6 ± 7.7 units. Figure 1 we and others demonstrated that also demonstrates the extravasation of Inflammation is recognized as a deletion of the gene encoding for eNOS FITC-dextran 70 through a series of fundamental underlying alteration in prevents mounting an appropriate images captured by intravital many diseases. One of the hallmarks of hyperpermeability response to VEGF microscopy and computer-assisted inflammatory processes is an increase (vascular endothelial growth factor) in digital image analysis. in microvascular permeability skin (Fukumura et al. 2001) and to PAF (hyperpermeability) to macromolecules. (platelet-activating factor) in cremaster Importantly, we demonstrated that An important function of muscle and in mesentery (Hatakeyama eNOS was the relevant nitric oxide hyperpermeability is to allow the et al. 2006). Figure 1 illustrates the synthase in this NO-regulated response movement of macromolecules for tissue dramatic difference in the PAF-induced as we established that mice lacking the healing and remodelling in response to hyperpermeability response in mouse gene encoding for inducible NOS injurious stimuli. Under inflammatory cremaster. Changes in microvascular (iNOS) produced a hyperpermeability conditions, microvascular permeability permeability were evaluated in the to PAF that was of comparable to macromolecules is controlled mainly interstitial space by integrated optical magnitude to that of wild-type mice at the postcapillary venules. The coordination of appropriate physiologic responses to the changing tissue environment in vivo is coordinated through signalling interactions between blood cells, vascular wall and parenchymal cells.

Nitric oxide is recognized as an important signalling regulator of cardiovascular function, but its role in the control of microvascular permeability was regarded as controversial. Evidence in tissues and in isolated venules demonstrated that the activity of endothelial NO synthase (eNOS) increases microvascular permeability to macromolecules in response to inflammatory agents (Ramírez et al. 1995; Yuan et al. 1993). Other results indicated that NOS activity prevents increases in permeability (Kurose et al. 1995). In either view, the evidence for Figure 1. Deletion of the gene encoding for eNOS (eNOS-/-) prevents PAF-induced hyperpermeability in mouse endogenous NO involvement as a cremaster muscle. Top panel shows changes in microvascular permeability using integrated optical intensity (IOI), expressed in arbitrary units, as an index. Bottom panel shows images of PAF induced changes in positive or negative modulator of permeability in cremaster muscles of wild-type and eNOS -/- mice (modified from J Physiol 574, 275-281, with permeability was persuasive but not permission).

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(Hatakeyama et al. 2006). We also showed that inhibition of eNOS by caveolin-1 scaffolding domain (an endogenous cellular regulator of eNOS activity) reduced the ability of PAF to increase microvascular permeability in cremaster muscle and mesentery of wild-type mice (Hatakeyama et al. 2006). The clinical significance of eNOS-derived NO is manifested by the reduction in postischemic microvascular permeability observed in cremaster muscle of eNOS-/- mice relative to wild-type mice (unpublished data) and by the increase in pulmonary microvascular permeability in caveolin­ 1 deficient mice (Schubert et al. 2002). Several laboratories, in further support of regulation of microvascular permeability by eNOS-derived NO, have documented that inhibition of eNOS decreases transport of macromolecules across monolayers of Figure 2. Simplified diagram of eNOS translocation hypothesis. eNOS is shown anchored (through endothelial cells, whereas substances myristoylation and palmitoylation) in caveolae and associated with caveolin-1 (purple arrows) in baseline conditions in endothelial cells. Upon stimulation by agonists, eNOS becomes phosphorylated, it dissociates from that phosphorylate eNOS and enhance caveolin-1 and translocates from the caveolae to either Golgi or cytosol. It is postulated that translocation to the NO production increase it (Lal et al. Golgi or cytosol compartments is related to vascular function and may allow eNOS to interact with its specific 2001; Yuan, 2000). function-related target proteins.

Mechanisms of signal may represent novel mechanisms or differential translocation of eNOS is transduction in pathways (Dudzinski et al. 2006). associated to microvascular function. hyperpermeability Using agonists that afford a relatively Much remains to be learned about how Enzyme location is of importance to clear discrimination between eNOS-derived NO signals for achieve its specific function. precapillary and postcapillary functions permeability regulation in Endothelial nitric oxide synthase is (basically vasodilation versus microvascular endothelia. While the found mainly in the cell membrane, but permeability), we observed in cells in gene sequence of eNOS is highly it is also well represented in Golgi culture that acetylcholine (a ‘pure’ conserved among species, the post­ under baseline conditions. The vasodilator) induces preferential translational regulatory modalities may relationship between eNOS location translocation to Golgi, whereas PAF (a vary in different species. A potentially and function might be of utmost hyperpermeability inducing agent) important regulatory mechanism relevance in endothelium due to the caused translocation preferentially to reported early is the phosphorylation of high NO-scavenging capacity of blood the cytosol (Sánchez et al. 2006). The eNOS by Akt (Fulton et al. 1999). haemoglobin. Translocation of eNOS basis for the physiological translocation However, recent data raise questions as from membrane to other cell of eNOS in vivo has been demonstrated to the specificity conferred to the signal compartments in response to agonists elegantly in transgenic mice using a by Akt-induced phosphorylation, has been documented by several fluorescent label that allows tracking particularly at Serine-1179. The techniques including conventional and the enzyme location in vivo (Cheng et questions are based on the observation confocal fluorescence microscopy as al. 2005). that, while necessary, this well as western blotting. Cellular phosphorylation step is common to location of eNOS may represent a The biological or experimental basis for several different NO-associated mechanism that determines the rate of the earlier controversy regarding NO vascular functions – such as NO production under different regulation remains unknown, but it may permeability, vasodilation, physiological or experimental reside in species differences and in angiogenesis, and cell survival conditions. Whether translocation inherent limitations of the use of non­ (Sánchez et al. 2006). The question is represents a defined mechanism or a specific pharmacologic agents. fundamentally valid even though step in signalling to bring the enzyme Compelling evidence based on the use several eNOS phosphorylation sites closer to its functional target remains to of genetically engineered mice have been described along with be determined. The diagram in Fig. 2 demonstrates that eNOS-derived NO regulation of the enzyme by protein­ describes a simplified view of regulates the necessary protein interactions, some of which experimental evidence indicating that hyperpermeability in postischemic

Physiology News | No. 65 | Winter 2006 | www.physoc.org FEATURES PN 23 tissues and in inflammation. Despite abundant and sophisticated information Acute pain obtained in cell cultures, the door is wide open for studies to elucidate the mechanisms by which eNOS-derived Pain is a perceptual experience that is NO exactly and specifically regulates ascribed to a body part and has a microvascular permeability in vivo. negative effect. Under normal circumstances, the stimuli that elicit Acknowledgments pain are tissue-damaging or would Our work was supported by NIH grant cause tissue damage were they to 5HL70634. persist: in short, noxious stimuli. Under these same conditions, noxious Peggy Mason Walter N Durán stimuli elicit a report of pain in verbal Takuya Hatakeyama* humans. Fabiola A Sánchez elicited by extreme pain is clearly to an Department of Pharmacology & Physiology, New While the gold standard for pain individual’s disadvantage, rendering her Jersey Medical School, University of Medicine and perception will always be verbal report, or him vulnerable to predators or Dentistry of New Jersey, Newark, NJ, USA and ontological and phylogenetic continuity potentially injurious situations. One *Department of Surgery, Seisho-kai Memorial Hospital, are most consistent with the idea that may postulate that while failing to Tokyo, Japan non-verbal humans and animals, at least serve an individual’s immediate safety mammals, experience pain. I will needs, painful stimulus-evoked References therefore use the terms pain and painful immobility is necessary to allow Cheng C, van Haperen R, de Waard M, van Damme LCA, Tempel D, stimulus to refer to what others term healing from an injury. According to Hanemaaijer L, van Cappellen GWA, Bos J, Slager CJ, Duncker DJ, van der Steen AFW, de Crom R, & Krams R (2005). Shear stress nociception and noxious stimulus. A this idea, if one moved about with a affects the intracellular distribution of eNOS: direct demonstration by fundamental distinction can be drawn ruptured disk, the disk would never a novel in vivo technique. Blood 106, 3691-3698. between acute pain – a transient heal, a reasonable possibility supported Dudzinski DM, Igarashi J, Greif D & Michel T (2006). The regulation experience – and persistent or chronic by the prevalence of prescribed bed rest and pharmacology of endothelial nitric oxide synthase. Annu Rev Pharmacol Toxicol 46, 235-276. pain. Acute pain requires a stimulus to for myriad painful conditions. Thus engage nociceptive pathways. In many being frozen in pain may be the only Fukumura D, Gohongi T, Kadambi A, Izumi Y, Ang J, Yun CO, Buerk DG, Huang PL & Jain R K (2001). Predominant role of endothelial forms of chronic pain, no stimulus long-term solution to injury despite its nitric oxide synthase in vascular endothelial growth factor-induced occurs and nociceptive pathways are immediate inconveniences. Finally, it angiogenesis and vascular permeability. Proc Natl Acad Sci U S A 98, 2604-2609. activated pathologically. should be noted that prolonged immobility is survivable only by Fulton D, Gratton JP, McCabe T J, Fontana J, Fujio Y, Walsh K, humans and other highly social animals Franke TF, Papapetropoulos A & Sessa WC (1999). Regulation of Acute pain is a common experience of endothelium-derived nitric oxide production by the protein kinase Akt. varied intensity, location and meaning. and is likely lethal in less social Nature 399, 597-601. Stubbing a toe or being scratched by a species. Hatakeyama T, Pappas PJ, Hobson II RW, Boric MP, Sessa WC & loving pet is transiently bothersome but Durán WN (2006). Endothelial nitric oxide synthase regulates rapidly forgotten. In contrast, breaking In addition to evoking either movement microvascular hyperpermeability in vivo, J Physiol (London) 574, 275-281. a toe or scratching a cornea evokes or active immobility, painful intense pain and the circumstances stimulation can also be ignored. This Lal BK, Varma S, Pappas PJ, Hobson RW 2nd & Durán WN (2001). VEGF increases permeability of the endothelial cell monolayer by surrounding such events are long happens when animals are feeding, activation of PKB/akt, endothelial nitric-oxide synthase, and MAP remembered. The acute pain voiding, or highly stressed (Baez et al. kinase pathways. Microvasc Res 62, 252-262. experience is at turns protective and 2005; Foo & Mason, 2005). For Kurose I, Wolf R, Grisham MB, Aw TY, Specian RD & Granger DN debilitating. Short-latency motor example, a rat that is eating chow will (1995). Microvascular responses to inhibition of nitric oxide production: role of active oxidants. Circ Res 76, 30-39. reactions to superficial pain stimuli, continue eating chow as heat is applied commonly withdrawals, are protective to his hindpaw, typically never Ramírez MM, Quardt SM, Kim D, Oshiro H, Minnicozzi M & Durán WN (1995). Platelet-activating factor modulates microvascular as are the longer latency, affectively responding to the heat stimulus that permeability through nitric oxide synthesis. Microvasc Res 50, 223­ motivated escape strategies including causes a brisk withdrawal when the rat 234. flight and freezing. Although freezing is not eating. While one may assume Sánchez FA, Savalia NB, Durán RG, Lal BK, Boric MP & Durán WN may not commonly be thought of as an that extremely intense noxious stimuli (2006). Functional significance of differential eNOS translocation. Am J Physiol:Heart & Circ Physiol 291, H1058-H1064. escape reaction, it is preferred to flight would elicit a reaction under all when a rodent faces a predator within circumstances, ethical concerns Schubert W, Frank PG, Woodman SE, Hyogo H, Cohen DE, Chow CW & Lisanti MP (2002). Microvascular hyperpermeability in close quarters and serves the same preclude these experiments and caveolin-1 (-/-) knock-out mice. Treatment with a specific nitric-oxide function as flight does in more spacious therefore only mild to moderate synthase inhibitor, L-NAME, restores normal microvascular permeability in Cav-1 null mice. J Biol Chem 277, 40091-40098. conditions. intensity stimuli are used. Allowing for context-specific modulation of noxious Yuan Y, Granger HJ, Zawieja DC & Chilian WM (1993). Histamine Pain can also be arresting, as anyone input allows for behavioral flexibility. increases venular permeability via a phospholipase C-NO synthase­ guanylate cascade. Am J Physiol 264, H1734-H1739. that has experienced an acute episode of back pain knows. Immobility (an Yuan SY (2000). Signal transduction pathways in enhanced Furthermore, this flexibility can serve microvascular permeability. Microcirc 7, 395-403. active state with high muscle tone) to adjudicate between the needs of an

Physiology News | No. 65 | Winter 2006 | www.physoc.org 24 PN FEATURES organism. For example, a rat that has had difficulty finding food may continue to eat, notwithstanding intense noxious stimulation. In contrast that same intense noxious stimulation will likely interrupt a rat that has recently fed. Under natural circumstances, food is relatively scarce and the system may be biased toward feeding, resulting in the failure of moderately painful stimuli to interrupt feeding.

Within the category of acute pain, there are significant differences and few similarities between cutaneous and deep pain. A noxious cutaneous stimulus typically leads to active movements away from the offending situation accompanied by multiple signs of sympatho-excitation. The resulting pain perception is well demarcated in time and space and if of Figure 1. A glaring example of the subtle effects of deep injury. A nail gun backfired, sending a nail into the cheek sufficient intensity, a long term memory of the young man operating the gun. Only a week later, after presenting with a toothache, did it become clear that of the event will ensue. In contrast, a the nail had actually entered the young man’s cranium and was lodged therein (photograph courtesy of the Littleton Adventist Hospital, Colorado, USA). noxious deep stimulus does not elicit movement and often is not immediately accompanied by perception, thereby phenomenologically, leading Thomas Taking the differences in behaviour and precluding explicit memory formation Lewis, an early pioneer in pain neural circuitry together leads to the of the surrounding circumstances. research, to lament that the somatic Gertrude Stein-esque conclusion that When of sufficient intensity, deep experiences of deep and superficial ‘acute pain is not acute pain is not injury can lead to perception, albeit pain share a common moniker (Lewis, acute pain’. Put more positively, acute poorly localized in time and space, as 1942). pain is heterogeneous, depending on well as immobility and sympatho­ location, intensity, and context. inhibition. Several differences exist between the neural mechanisms that support acute Peggy Mason Several glaring examples of the subtle cutaneous and deep pain. Department of Neurobiology, Pharmacology and Physiology, The University of Chicago, Chicago, IL, effects of deep injury have recently USA been reported. In one such example, a Afferents carrying information about nail gun backfired, sending a nail into cutaneous pain belong to a specific References the cheek of the young man operating class of nociceptors that are activated only by noxious stimuli that would Baez MA, Brink TS & Mason P (2005). Roles for pain modulatory the gun. After receiving treatment for cells during micturition and continence. J Neurosci 25, 384-394. the cutaneous wound in his cheek, the normally elicit pain. Afferents that man returned home. Only a week later, carry deep pain input are more Cervero F, Lumb BM & Tattersall JE (1985). Supraspinal loops that mediate visceral inputs to thoracic spinal cord neurones in the cat: after presenting with a toothache, did it heterogeneous with some coding pain involvement of descending pathways from raphe and reticular become clear that the nail had actually exclusively, like cutaneous nociceptors, formation. Neurosci Lett 56, 189-194. entered the young man’s cranium and and some that code both pain and non­ Foo H & Mason P (2005). Sensory suppression during feeding. Proc was lodged therein (see Fig. 1). painful information. Natl Acad Sci U S A 102, 16865-16869. Perhaps even more remarkable was the Ness TJ & Gebhart GF (1988). Colorectal distension as a noxious fact that this was the second such case Another difference between cutaneous visceral stimulus: physiologic and pharmacologic characterization of pseudaffective reflexes in the rat. Brain Res 450, 153-169. observed at the suburban Denver and deep pain is the role of descending hospital where the injury was treated. It modulation from the brainstem and Lewis T (1942). Pain. The Macmillan Company, New York. is telling to contrast the immediate forebrain. The net effect of descending indifference evoked by such a severe modulation onto cutaneous nociceptive The Third Meeting on pathways is a tonic inhibition. In deep tissue injury as described above Molecular mechanisms of marked contrast, it appears that for with the inordinate concern and neurodegeneration visceral nociception, a spino-bulbo­ awareness that accompanies a will take place in Milan, Italy spinal loop is required for expression superficial injury such as a paper cut. from 19-21 May 2007 of the full behavioral and physiological Full details at In sum, it is clear that cutaneous and reaction (Cervero et al. 1985; Ness & deep pain are very different Gebhart, 1988). http://users.unimi.it/triplet/2007

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Legs pay out for cost of breathing! Lee Romer and Jerome Dempsey propose that the demand placed on the respiratory muscles during high-intensity endurance exercise plays a pivotal role in determining exercise performance through its effect on limb blood flow and muscle fatigue

The respiratory muscles are, with few diaphragmatic work mean large shares exceptions, structurally suited to the of the total cardiac output are devoted high ventilatory demands of endurance to the diaphragm, whereas during exercise. The diaphragm, for example, exercise the diaphragm must compete has a high oxidative capacity, multiple with locomotor muscles for their share sources of blood supply and a unique of the available cardiac output. Less resistance to vasoconstrictor influences, blood flow to the diaphragm promotes making it the most fatigue resistant of inadequate oxygen transport and all skeletal muscles. Such structural increases the likelihood of fatigue. characteristics, in combination with precise neural regulation of breathing The aforementioned physiological pattern and respiratory muscle effects of the work of breathing have recruitment, mean that the capacity of important functional consequences, as these muscles for pressure generation is demonstrated by an approximately 15% usually well in excess of the demands improvement in performance during placed upon them. Thus, in most high-intensity endurance exercise with untrained healthy individuals the partial unloading of the respiratory pressures produced by the inspiratory muscles via mechanical ventilation muscles during progressively increasing (Harms et al. 2000). One aspect of exercise are only about 50% of Jerome Dempsey (left) and Lee Romer respiratory muscle work that might maximum dynamic capacity, the limit exercise performance is a reflex oxygen cost of breathing approximates diaphragmatic work incurred during effect from fatiguing respiratory 10% of maximum oxygen uptake and exercise. Indeed, fatigue does not muscles, which increases sympathetic diaphragm fatigue does not occur. occur until diaphragmatic work is vasoconstrictor outflow and voluntarily increased two-fold greater compromises perfusion of limb muscle Despite the considerable aerobic than that required during maximal thereby limiting its ability to perform capacity of the diaphragm, this muscle exercise. The probable explanation of work. Fatiguing contractions and will fatigue in response to exercise this substantial effect of whole body accumulation of metabolites in the sustained to exhaustion at intensities exercise on diaphragm fatigability is respiratory muscles activate the greater than 80-85% of maximum that, at rest, the volitional increases in unmylenated Type IV phrenic afferents oxygen uptake. For example, the pressure measured across the diaphragm in response to bilateral stimulation of the phrenic nerves (1-20 Hz) was reduced 15-50% below pre­ exercise baseline and did not return to control values until 1-2 hours after exercise (Johnson et al. 1993). The reason for this exercise-induced diaphragmatic fatigue is due in part to the high levels of diaphragmatic work that must be sustained throughout high­ intensity exercise, as demonstrated by the finding that diaphragmatic fatigue was prevented when diaphragmatic work during exercise was reduced by over 50% using a mechanical ventilator. However, other factors besides diaphragmatic work must also be responsible for the exercise-induced Figure 1. Schematic of the proposed respiratory muscle metaboreflex and its effects. The metaboreflex is diaphragmatic fatigue, because fatigue initiated by fatigue of the inspiratory muscles, mediated supra-spinally via phrenic afferents, leading to sympathetically mediated vasoconstriction of limb locomotor muscle vasculatures, exacerbating peripheral fatigue does not occur when the resting subject of working limb muscles and (via feedback) intensifying effort perceptions, thereby contributing to limitation of mimics the magnitude and duration of high-intensity, endurance exercise performance.

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of unloading on fatigue represents an underestimation of what might be attributed to the total work of breathing. For example, the normal work of breathing was reduced by only slightly more than one half. Furthermore, the loading and unloading arms of the study were confined to inspiration. Recent evidence in healthy individuals suggests that the expiratory abdominal muscles also fatigue in response to high-intensity endurance exercise and that expiratory loading to the point of task failure elicits an increase in leg muscle sympathetic nerve activity. Furthermore, the high expiratory pressures often encountered during high-intensity exercise in response to active expiration and expiratory flow Figure 2. Stimulation of the femoral nerve using a double coil connected to two magnetic stimulators. limitation have been shown to reduce Quadriceps muscle force output was assessed using a force transducer connected to a strap around the subject’s stroke volume, which may be expected right ankle. The potentials (M-waves) evoked from the quadriceps via nerve stimulation were measured using surface electromyography. to exacerbate limb fatigue via concomitant reductions in limb blood which, via a supraspinal reflex, evokes constructed by altering the duration flow. an increase in muscle sympathetic between paired twitches. These nerve activity (St Croix et al. 2000) and measurements were performed before In health, it is likely that exercise­ a reduction in blood flow and vascular and after high-intensity cycle exercise induced respiratory muscle work conductance in the resting leg (Sheel et under conditions of control, inspiratory contributes to limb fatigue only during al. 2001) (see Fig. 1). During maximal muscle unloading (via mechanical sustained high-intensity exercise that is exercise, concomitant increases in ventilation) and inspiratory muscle sufficient to elicit significant levels of vascular conductance and blood flow to loading (via inspiratory resistors). fatigue in the diaphragm or accessory working limb muscles have been When exercise at 90% of peak work respiratory muscles. Environmental observed when the normally occurring rate was continued to exhaustion with hypoxia, and particularly chronic work of breathing is reduced using a breathing unimpeded, quadriceps twitch hypoxic exposure which potentiates the mechanical ventilator (Harms et al. force averaged across a wide range of hyperventilatory response to exercise 1997). Furthermore, in resting and stimulation frequencies (1-100 Hz) was and consequently the work of exercising dogs, a bolus injection of reduced by almost 30% in the period breathing, might also be expected to lactic acid into the diaphragm by way immediately after exercise. When the exacerbate the effect of respiratory of the phrenic artery elicits a transient exercise was repeated on a different day muscle work on limb fatigue. From a reduction in hind limb blood flow and at equal workload and duration as the clinical perspective, changes in the vascular conductance – these changes control condition, but with the normally work of breathing may play a appear to be mediated sympathetically occurring work of breathing reduced by particularly important role in because they are prevented by 50-60%, the decrease in quadriceps determining limb fatigue in patients pharmacological blockade of the force output was attenuated by about with chronic obstructive pulmonary adrenergic receptors (Rodman et al. one-third. Increasing the work of disease (COPD) and chronic heart 2003). breathing by 80% above control failure (CHF). In patients with COPD exacerbated the severity of fatigue by the limb muscles are more fatigable In our most recent experiments we about 40% compared with identical than in healthy subjects and the attempted to determine whether the exercise with breathing unimpeded. inspiratory and expiratory pressures aforementioned changes in vascular developed during exercise are conductance and limb blood flow with The effect of inspiratory muscle substantial. Many such patients exhibit manipulation of the work of breathing unloading on limb muscle fatigue was a plateau in leg blood flow early during impact upon the fatigability of the consistent among subjects but relatively progressively increasing exercise and working limb muscles (Romer et al. small compared with the much larger these patients also tend to have the 2006). We assessed limb muscle reductions in limb fatigue we recently highest levels of ventilation and fatigue by magnetically stimulating the found by preventing the mild arterial dyspnea during exercise, suggesting femoral nerve using a paired twitch oxygen desaturation that can that blood flow may be redirected to technique (see Fig. 2). Force:frequency accompany high-intensity exercise. the respiratory muscles. Patients with curves for the quadriceps were Nevertheless, it is likely that this effect CHF also have an elevated work of

Physiology News | No. 65 | Winter 2006 | www.physoc.org FEATURES PN 27 breathing, in addition to blunted cardiac Functional domains of tropomyosin: new insights output and sympathetic vasoconstrictor responses to exercise. In combination, Functional studies of tropomyosin molecule using these factors could compromise limb transgenic approaches reveal that its inner core region has blood flow and fatigue, even during a greater effect on calcium sensitivity of the myofilaments submaximal exercise. and the carboxy terminal ends regulate the TM-actin affinity In summary, unloading the respiratory and/or crossbridge kinetics muscles in normal healthy individuals improves performance during high­ Tropomyosin (TM) is a thin filament intensity exercise coincident with regulatory protein that is intricately increases in limb vascular conductance involved in cardiac muscle contraction. and blood flow and a reduction in leg Working in conjunction with the muscle fatigue. We propose that troponin complex, TM’s position on exercise-induced respiratory muscle actin either blocks or exposes myosin­ work contributes to both ‘peripheral’ binding sites, and thus prevents or and ‘central’ fatigue influences on allows crossbridge formation as exercise performance. In other words, outlined in the three state model of quadriceps muscle fatigue plays a muscle contraction (McKillop & pivotal role in determining exercise Geeves, 1993). Mariappan Muthuchamy (left) and Robert Gaffin performance both through its direct effect on muscle force output (i.e., The principal focus of our lab has been α-TMHis276Asn TG mice did not alter peripheral fatigue) and also through its to address the role of TM in cardiac calcium sensitivity but decreased -dP/dt feedback effect on effort perceptions, muscle function. Initial findings using (Gaffin et al. 2004b). Furthermore, TG causing reduced motor output to the β-TM transgenic (TG) mouse model mouse lines that express a mutant form working limb muscles (i.e., central demonstrated that β-TM TG hearts of α-TM in which the first 275 residues fatigue). displayed diastolic dysfunction are from α-TM and the last nine amino including increased time to one-half acids are from β-TM (α-TM9aa∆β) Acknowledgements relaxation and a decreased maximum exhibit decreased rates of contraction The authors are grateful to the NHLBI rate of relaxation in working heart and relaxation in working heart studies for supporting their work. preparations, increased calcium (Gaffin et al. 2006). The myofilaments sensitivity in skinned fibres. containing α-TM9aa∆β protein Lee M Romer1 demonstrate a normal pCa-force Jerome A Dempsey2 Two important charge modifications in relationship (Gaffin & Muthuchamy, 1 β-TM, Ser229Glu and His276Asn, give unpublished data). These data indicate Centre for Sports Medicine and Human Performance, α Brunel University, Uxbridge, UK it a charge that is more negative than - that the function of TM is 2John Rankin Laboratory of Pulmonary Medicine, TM. Since the remaining residue compartmentalized along its length Department of Population Health Sciences, University differences between the two isoforms (Fig. 1). of Wisconsin, Madison, USA involve rather conservative polar or non-polar changes, we hypothesized We propose that residues located near References that generating β-TM’s two charge Harms CA, Babcock MA, McClaran SR, Pegelow DR, Nickele GA, the site where TM interacts with the Nelson WB & Dempsey JA (1997). Respiratory muscle work modifications on an α-TM background C-terminus of TnT (which also interacts compromises leg blood flow during maximal exercise. J Appl Physiol would produce results identical to those 82, 1573-1583. with the C-terminus of troponin C seen in β-TM TG mice. Thus, we (TnC), the myofilament’s calcium Harms CA, Wetter TJ, St Croix CM, Pegelow DF & Dempsey JA generated ‘double mutation’ sensor) primarily affect calcium (2000). Effects of respiratory muscle work on exercise performance. J Appl Physiol 89, 131-138. (Ser229Glu and His276Gln) TG mouse sensitivity since α-TMSer229Glu lines. These TG mice exhibited caused a decrease while Johnson BD, Babcock MA, Suman OE & Dempsey JA (1993). 2+ Exercise-induced diaphragmatic fatigue in healthy humans. J Physiol decreased Ca -sensitivity in skinned α-TMHis276Asn and α-TM9aa∆β had 460, 385-405. fibres as well as a decrease in both the no effect. On the other hand, Rodman JR, Henderson KS, Smith CA & Dempsey JA (2003). rate of contraction and relaxation in α-TMHis276Asn and α-TM9aa∆β Cardiovascular effects of the respiratory muscle metaboreflexes in working heart preparations (Gaffin et mutations located in the carboxy dogs: rest and exercise. J Appl Physiol 95, 1159-1169. al. 2004a). terminus of TM, have altered the rates Romer LM, Lovering AT, Haverkamp HC, Pegelow DF & Dempsey of contractile dynamics without JA (2006). Effect of inspiratory muscle work on peripheral fatigue of locomotor muscles in healthy humans. J Physiol 571, 425-439. We then wished to see the effects of affecting calcium sensitivity. these two charge changes on an Sheel AW, Derchak PA, Morgan BJ, Pegelow DF, Jacques AJ & α Dempsey JA (2001). Fatiguing inspiratory muscle work causes reflex individual basis. -TMSer229Glu TG To explain this, we point to the fact that reduction in resting leg blood flow in humans. J Physiol 537, 277­ mice exhibited a decrease in the carboxy terminus affects TM-actin 289. myofilament calcium sensitivity and a affinity in the ‘open’ state of muscle St Croix CM, Morgan BJ, Wetter TJ & Dempsey JA (2000). Fatiguing decrease in both +dP/dt and -dP/dt in contraction as proposed by Hitchcock­ inspiratory muscle work causes reflex sympathetic activation in humans. J Physiol 529, 493-504. the working heart while Degregori’s group (Moraczewska et al.

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on calcium sensitivity due to their proximity to the TnT-TnC interface while residues in the carboxy terminus affect either TM-actin affinity or crossbridge kinetics. Furthermore, these studies support the idea that the localised flexibility present in the coiled coil structure of various TM isoforms are different, and that plays an important role in interacting with neighboring thin filament regulatory proteins and differentially modulating the myofilament activation processes.

Acknowledgements This work was supported by an NIH grant HL-60758 to M M.

Robert D Gaffin Figure 1. Compartmentalisation of TM’s function along its length. Schematic representation of tropomyosin (TM) Mariappan Muthuchamy and troponin complex interaction is shown. In the inner core region of tropomyosin (TM), the C-terminus of troponin T (TnT) binds to TM as well as to the C-terminus of both TnC and the N-terminus of troponin I (TnI). At Cardiovascular Research Institute and Department of the overlap region of TM, the N-terminus of TnT interacts. Residues near the inner core region of TM primarily Systems Biology and Translational Medicine, Texas affect calcium sensitivity due to the proximity of the myofilament’s calcium sensor, TnC. Residues in the C­ A&M Health Science Center, TX, USA. terminus of TM primarily affect TM-actin affinity and/or crossbridge kinetics. References 1999). An enhancement of TM-actin contractile dynamic changes and/or Gaffin RD, Gokulan K, Sacchettini JC, Hewett T, Klevitsky R, affinity during this phase would delay calcium sensitization of the Robbins J & Muthuchamy M (2004a). Charged residue changes in the carboxy-terminus of alpha-tropomyosin alter mouse cardiac the myofilament’s transition from the myofilaments. However, none of these muscle contractility. J Physiol 556, 531-543. ‘open’ to the ‘blocked’ state thus affect changes, either individually or in Gaffin RD, Gokulan K, Sacchettini JC, Hewett TE, Klevitsky R, contraction and relaxation kinetics in combination within the α-TM Robbins J, Sarin V, Zawieja DC, Meininger GA & Muthuchamy M α-TM His276Asn and α-TM9aa∆β TG molecule, mimic the function that is (2006). Changes in end-to-end interactions of tropomyosin affect mouse cardiac muscle dynamics. Am J Physiol Heart Circ Physiol mice. seen in the β-TM TG mice. In 291, H552-563. addition, these TG mice functional Compartmentalization of TM function Gaffin RD, Tong CW, Zawieja DC, Hewett TE, Klevitsky R, Robbins J studies reveal there is a lack of & Muthuchamy M (2004b). Charged residue alterations in the inner­ is further evidenced by the recent work correlation between the force-calcium core domain and carboxy-terminus of alpha-tropomyosin differentially affect mouse cardiac muscle contractility. J Physiol 561, 777-791. of Dr Wieczorek’s group. TG mice relationships and the haemodynamic expressing chimeric TM protein (exons measurements at the myofilaments and Jagatheesan G, Rajan S, Petrashevskaya N, Schwartz A, Boivin G, α Arteaga G, de Tombe PP, Solaro RJ & Wieczorek DF (2004). 1–8 [residues 1– 257] from -TM plus the whole heart levels, respectively, β Physiological significance of troponin T binding domains in striated exon 9 [residues 258–284] from -TM) indicating a cautious extrapolation/ muscle tropomyosin. Am J Physiol Heart Circ Physiol 287, H1484­ 1494. exhibit decreased rates of contraction correlation of the physiological and and relaxation in whole heart studies, biochemical parameters of cardiac Jagatheesan G, Rajan S, Petrashevskaya N, Schwartz A, Boivin G, and a decrease in calcium sensitivity in Vahebi S, DeTombe P, Solaro RJ, Labitzke E, Hilliard G & Wieczorek function. On the other hand, both DF (2003). Functional importance of the carboxyl-terminal region of skinned fibres (Jagatheesan et al. calcium activation of the thin filament striated muscle tropomyosin. J Biol Chem 278, 23204-23211. 2003). In addition, exchanging the TnT and positive feedback by strong α McKillop DF & Geeves MA (1993). Regulation of the interaction binding domains of -TM amino acids crossbridges mechanisms make between actin and myosin subfragment 1: evidence for three states β [175–190 and 258–284] with the -TM significant contributions to each phase of the thin filament. Biophys J 65, 693-701. regions decreases the rates of of the contractile cycle. Further Moraczewska J, Nicholson-Flynn K & Hitchcock-DeGregori SE contractile dynamics; however, the (1999). The ends of tropomyosin are major determinants of actin analytical tools to quantify the affinity and myosin subfragment 1-induced binding to F-actin in the calcium sensitivity is not altered contributions of the calcium or open state. Biochemistry 38, 15885-15892. (Jagatheesan et al. 2004). crossbridge cooperative effects to each phase of the contractile cycle would be These results, along with our data, useful to address the mechanisms of European Association for the Study of Obesity point out the complexity of the TM function in the myofilament th relations between structure and function activation processes. 15 European Congress on of TM molecule. Each difference exists Obesity between the α- and β-TM molecules In summary, we conclude that the Budapest, Hungary such as TnT binding domains, charge function of α-TM is compartmentalised 22-25 April 2007 residue changes or the changes in the along its length. Sequences nearer the carboxy terminal ends contribute to the inner core of TM have a greater effect http://www.eco2007.org

Physiology News | No. 65 | Winter 2006 | www.physoc.org KIEV INTERNATIONAL WORKSHOP PN 29

The study of nociception from periphery to brainstem An International Workshop at the Bogomoletz Institute for Physiology, Kiev, Ukraine, 4-7 June 2006

Taking an idea through to both oral and practical elements. fruition: organizing an Where will we accommodate everyone, International Workshop in how shall we keep them occupied for Kiev the scheduled 4 days of our workshop? For the organizers of any international What about that other most essential event, one of the fun parts is sitting ingredient – the social programme? We casually over a glass of wine and take our hats off to our co-organizers drafting a list of high profile Oleg Krishtal, Nana Voitenko, Olga participants to invite to your event. It Lyubanova and Sergei Khasabov. Their is not always the case that the ‘wish unbounded enthusiasm and support for list’ becomes reality but this certainly the workshop carried it forward and was the case for our Kiev-based student truly made it the success that it was. workshop. Perhaps it was the fact that They made it feasible to run a it was to be hosted in the eponymous workshop that delivered a series of 12 Bogomoletz Institute of Physiology plenary lectures, 10 demonstrations and which, under the leadership of Platon hands-on practicals, three poster Kostyuk, has contributed so much to sessions and a social programme that neuroscience, or maybe it was the generated a real feeling of camaraderie location in Kiev, one of Europe’s most amongst a group of people meeting, in architecturally rich and beautiful cities many cases, for the first time. situated on the Dnepr river, or perhaps it was the enticing social events – The plenaries covered topics ranging whatever, the fact remains that we were from the peripheral neural encoding of fortunate enough to assemble a first pain, through to spinal or brainstem circuitry, regulatory processing and the Top: Tombstone commemorating Oleg Bogomoletz, class programme of more than a dozen founder and original President of the Institute in 1953. speakers from across Europe and the challenge of analgesic drug discovery. Above: Student delegates Andre Zaharenko (Brazil, USA. But what about the student Practical sessions provided the students white t-shirt, then clockwise) Mino Belle (UK), Lianne delegates – will anyone actually attend with an insight to in vitro and in vivo Leith (UK), Nadia Cerminara (UK) Charlotte Flavell methodologies, electrophysiology, light (UK), Péter Szûcs (Hungary), Sergiy Lytvynenko our event? Again, we need not have (Ukraine), Aleksey Boldyryev (Ukraine), Volodymyr worried as the response to our call for and confocal microscopy, behavioural Cherkas (Ukraine), Stella Koutsikou (UK) gazing into applications was overwhelming and the physiology and molecular biology, as the future? quality of submitted cvs impressive. applied to pain research. Below, left: Participants and speakers assemble on the steps of the Bogomoletz Institute. After much deliberation, a total of 45 postgraduate and post-doctoral student In the poster sessions, the students were the best laboratory demonstration, as delegates were selected from Institutes able to present their own data and voted by the students. Interest beyond and Universities of UK, Ukraine, prizes were awarded to the best two the confines of the Bogomoletz was Romania, Slovakia, Russia, Italy, posters. We acknowledge gratefully the evidenced by an invitation to hold a Hungary, Bulgaria, Belarus, Czech support of Bill Willis (UTMB, Texas) televised press conference with local Republic, Georgia and Brazil. and Ray Hill (MSD, UK) who enabled us to provide as prizes much coveted media – questions on pain in the Now our thoughts turned to the copies of the new edition of Willis & context of ‘sado-masochism’ were side­ workshop schedule itself and the Coggeshall’s Sensory mechanisms of stepped as diplomatically as possible. practicalities of providing a stimulating the spinal cord. A book prize was also But what of the social events? The scientific programme that combined kindly donated by Platon Kostyuk for unforgettable ‘Walking tour of Kiev’ that tested the stamina of many a delegate, the boat trip and BBQ on the beautiful river Dnepr with optional swimming and impromptu fireside concert, the fabulous suppers in traditional settings washed down with capsaicin-laced vodka followed by music, singing and (limbo!) dancing.

Many things contributed to the success of this workshop but may we take this

Physiology News | No. 65 | Winter 2006 | www.physoc.org 30 PN KIEV INTERNATIONAL WORKSHOP opportunity to say a huge ‘thank you’ to the end of the event that our efforts the speakers for giving up their were not in vain and that all our goals valuable time to us, to all at were accomplished. The utterance of Bogomoletz for the resources that were Jürgen Sandkühler, one of our esteemed put at our disposal, to Physiological invited speakers, was among our Society staff, especially David Bennett rewards: ‘I have put Kiev in my list of who efficiently attended to all manner cities the invitation from where I will of requests before, during and after the never reject!’ event, and finally, but not least, to the student delegates whose enthusiasm Oleg Krishtal and energy for their science was Nana Voitenko Bogomoletz Institute of Physiology, Kiev tangible. Co-organizers Bridget M Lumb University of Bristol, UK Student delegate report Anne E King When I started my PhD, I had a lot of University of Leeds expectations about conferences. I Organisers thought of them as places where we could meet people working in the same The host organizers field; and ask them for help and advice. perspective However, after some years, I realized it Ukraine is gradually strengthening, competitive and internationally was just not that easy. Poster sessions, both socially and economically, after a recognized. Today, the Institute is a and discussions between lectures, are difficult period of political transition leading scientific research centre in just too short to meet everyone; and during the 1990s. Despite some Ukraine in the areas of molecular how can I find someone who I have progress, the financial climate for physiology, biophysics, read a lot about, but never seen? science and its future development neurophysiology and pathological Generally, they are not the poster remains austere, with government physiology. In the context of allowing presenters and they clip their badges on support still below perceived desirable us to showcase to both the Ukrainian their trousers where I cannot read their levels. Bogomoletz Institute of Government and the global scientific names, even with glasses. This Physiology in Kiev is one of only a few community our ability to conduct state­ workshop gave us the very opportunity academic institutions that has not only of-the-art research and to host to meet those scientists, who have the managed to survive, but maintains a international events, this workshop was largest reputation in the field of level of research that is scientifically extremely important. Without false nociception and pain research. We Below: Dmitrii Vasilenko (Ukraine) takes to the water modesty, we can state that the purpose could learn a lot from their talks, get a during the river boat trip and BBQ. of the workshop, which was to provide very good overview of different areas Centre, top:: Enjoying the reception supper, co­ step-by-step methods and protocols of of pain research, and see the theoretical organizers Oleg Krishtal and Nana Voitenko with multidisciplinary approaches related to background of different physiological Eugen Grishin (Russia). Centre, below: Co-organizers Sergei Khasabov (USA) the study of pain transduction, was techniques. Post-docs could learn how and Olga Lyubanova (Ukraine) take time out with Ray fulfilled. High attendance at all of the Hill (UK). seminars and practical demonstrations Far right, top: An impromptu sing-along sets the BBQ by students, who had travelled to Kiev alight. Far right, below: Organizers Anne King (UK) and from many world famous laboratories, Bridget Lumb (UK) enjoy a glass of wine with student indicated that the areas of research delegates Dinu Crenguta (Romania) and Gayane covered by our workshop were sound. Margaryan (Italy). The vibrancy of the student poster sessions and many discussions between delegates testified to the quality and topicality of data they presented. The activity of the scholars suggested that the practical demonstrations had been worthwhile – even for those who were familiar with modern methods and approaches. We would like to thank all the Institute researchers who spent their valuable time preparing practical demonstrations that were interesting for all of the students from different areas of pain research. It was a real pleasure for the host organizers to recognize at

Physiology News | No. 65 | Winter 2006 | www.physoc.org KIEV INTERNATIONAL WORKSHOP PN 31

Enigma of P2X3 receptors: how to reconcile their properties with putative role in the nociception? The receptors for ATP in sensory neurons are functional in

homomeric (P2X3) and heteromeric (P2X2/3) forms. Which of them could be more important for nociception?

When locally applied to the tissues, ATP causes pain. The molecular basis Student delegate Volodymyr Cherkas (Ukraine) for priming this sensation is provided sharpens up his limbo dancing skills. by at least two types of receptors that their professors thought about different open cationic (Na+ and Ca2+ permeable) topics and designed their experiments. channels when bound to ATP. These Time for formal discussions was short, receptors (P2X2 and P2X3, forming also but we had time enough to talk to heteromeric P2X2/3) are expressed in anybody during social events. Of the majority of sensory neurons course, I don’t mean this could happen belonging to dorsal root (DRG), Volodymyr Khmyz (left), Oleg Krishtal and Oleksandr at the end of a 3 hour walk in Kiev, trigeminal and nodose ganglia of Maximyuk when nobody had air in his/her lungs mammals. The release of ATP except for the lady who was the guide! apparently occurs when the tissue cells In The Bogomoletz Institute, we had a are destroyed or in response to In response to extracellular ATP, P2X3 chance to see a lot of physiological and deformation of hollow organs, when it receptors gate current that rapidly morphological techniques that are used is supposed to initiate visceral pain. declines upon its onset, supposedly in our field. Of course, 30-40 minutes reflecting desensitization. In the case of are far too short to learn a technique P2X3 receptors are widely expressed in sole activity of P2X3 receptors (rarely but, at least, we got an impression and the small nociceptive sensory neurons. observed pattern of response in the important information on who to ask if Their role in nociception has been somata of DRG neurons), the residual we want to do something similar. revealed in numerous studies including steady-state current is virtually absent. Everyone who demonstrated his/her knock-out experiments (Souslova et al. The removal of desensitization is own experiment was very enthusiastic 2000) and various pain models. It extremely slow. Various groups of and keen to do his/her best. We have seems to be especially prominent in investigators have reported that from had many nice social events, but my inflammation. Recently our group has 20-30 min are needed to obtain the next best memories are of the boat trip on found that P2X receptors in sensory response of similar amplitude. This the Dnepr river. During the trip we neurons and fibers are under opioid property of P2X3 receptors results in at enjoyed a spectacular view of Kiev and control (Chizhmakov et al. 2005). least two problems arising at different the boat docked on a small island, with untouched habitat. It was quite cold, so Dmitri Vasilenko’s swim in the Dnepr seemed a real miracle (I had two jumpers and a coat on!). While the Saslik (a grilled pork meat) was prepared, some guys made a fire, and that provided a very nice warm feeling. Ruslan Mazgutov had a guitar and started to play. It is time we thanked him for these moments. Many of the participants who knew these, mainly Russian, songs were singing along with him. It is hardly possible to describe the atmosphere which appeared in minutes and took me back to my teenage years. I would like to say many thanks for all the organisers and Figure 1. Response of a neuron acutely isolated from DRG of rat to externally applied ATP (100 µM, saturating participants who made this workshop concentration). unforgettable for many of us. A, response elicited by a short (6 ms) application of ATP (red trace) is identical to the response elicited by a prolonged application of the agonist (black trace). B, the response recorded in 2 s after the short application of Zita Puskar ATP is completely desensitized (green trace). The response recorded in 10 min after a short application (red Semmelweis University, Budapest, Hungary trace) is still considerably desensitized, though it is larger than the response measured in 10 min after a long Delegate application (black trace).

Physiology News | No. 65 | Winter 2006 | www.physoc.org 32 PN KIEV INTERNATIONAL WORKSHOP levels: of this site to ATP is high: use­ middle-size DRG neurons acquire

• it is difficult to reconcile the dependent desensitization occurs in robust rapid (P2X3 receptor-mediated) desensitization removal being so slow nanomolar range of background ATP response to ATP. Its functional role at with their putative role in concentration. the periphery may well be other than nociception, and the nociception. • it is just difficult to study the function Thus, when bound to ATP, P2X3 of these receptors. receptor is prone to activation and Acknowledgements desensitization with a subsequent ultra­ Research in the authors’ laboratory is The latter problem became obvious slow removal of the latter occurring via supported by grants from CRDF and immediately after our lab discovered unbinding of ATP. Our experiments HHMI. the ATP-sensitivity of sensory neurons: with rapid ATP removal seem to Volodymyr Khmyz we tried to overcome it by removing confirm this picture in its main details. Oleksandr Maximyuk ATP from the membrane before it Fig. 1B demonstrates that Oleg Krishtal would cause significant desensitization desensitization elicited by short (‘square-pulse’ application technique Department of Cellular Membranology, Bogomoletz application of ATP lasts for many Institute of Physiology, Kiev, Ukraine (Kryshtal et al. 1982)). However, this minutes. Desensitization removal takes approach, successful in eliciting 15-20 min in the case of short References perfectly reproducible responses of application vs. 20-30 min for a long Chizhmakov I, Yudin Y, Mamenko N, Prudnikov I, Tamarova Z & much slower P2X2 and P2X2/3 one. This slight difference may indicate Krishtal O (2005). Opioids inhibit purinergic nociceptors in the sensory neurons and fibres of rat via a G protein-dependent receptors, gave only partial relief in a possible existence of several mechanism. Neuropharmacology 48, 639-647. case of the most rapid P2X3 receptors: desensitized states. Cockayne DA Hamilton SG, Zhu QM, Dunn PM, Zhong Y, Novakovic it was impossible to make the square S, Malmberg AB, Cain G, Berson A, Kassotakis L, Hedley L, Lachnit pulse of agonist application much Thus, our data suggest that the ‘duty WG, Burnstock G, McMahon SB & Ford AP (2000). Urinary bladder hyporeflexia and reduced pain-related behaviour in P2X3-deficient shorter than 100 ms and this time was cycle’ of an open channel gated by mice. Nature 407, 1011-1015. already sufficient to result in nearly P2X3 receptor negligibly depends on Kryshtal' OA, Marchenko SM & Pidoplichko VI (1982). [ATP receptor complete decay of the response. the presence of agonist after the latter in the membrane of sensory neurons]. Dokl Akad Nauk SSSR 267, Recently we returned to this problem has been bound by the receptor. 1244-1247. by using piezoelectric drive of the Unfortunately we still know nearly Pratt EB, Brink TS, Bergson P, Voig MM & Cook SP (2005). Use- microfilaments used for the application nothing about the temporal pattern of Dependent Inhibition of P2X3 Receptors by Nanomolar Agonist. of ATP to the cell. This technique, ATP release/removal which occurs at Journal of Neuroscience 25, 7359-7365. introduced first by Dudel and the ATP-sensitive nerve endings in vivo. Souslova V, Cesare P, Din Y, Akopian AN, Stanfa L, Suzuki R, Carpenter K, Dickenson A, Boyce S, Hill R, Nebenuis-Oosthuizen D, colleagues, enabled us to apply ATP to Smith AJ, Kidd EJ & Wood JN (2000). Warm-coding deficits and the cell for the time as short as 4-7 ms Activation of P2X3 receptors develops aberrant inflammatory pain in mice lacking P2X3 receptors. Nature (depending on the cell size), washing in the range of micromolar 407, 1015-1017. out the ligand just at the peak of concentrations of the agonist, while response. We have found a remarkable their desensitization occurs at International Workshop picture: the kinetics of current decay nanomolar concentrations. The Kiev appear to be completely independent on background concentration of ATP in the Invited speakers tissues varies within this range of the presence of agonist at the Eugene V Grishin (Moscow, Russia) membrane (Fig. 1A). This could values in a function-dependent manner, indicate that the rapid decay of the reaching micromoles in the working Ray Hill (MSD, UK) response of P2X3 receptors to ATP muscles. Designed as they are, P2X3 Steve Hunt (London, UK) reflects deactivation, not receptors should be sensitive to rapid Sergey Khasabov (Minnesota, USA) desensitization. However, subsequent acts of significant ATP release and Platon Kostyuk (Kiev, Ukraine) trial (in 2s after a short application of remain silent long time after being Alexander Kostyukov (Kiev, Ukraine) the agonist) demonstrates that the activated. It remains enigmatic, how to desensitization is complete (Fig. 1B). reconcile this property of P2X3 Deolinda Lima (Oporto, Portugal) Correspondingly, our finding better fits receptors with their putative function as Jiri Palacek (Prague, Czech Republic) another picture. Recently it has been nociceptors. Taking into account their Miklos Rethelyi (Budapest, Hungary) demonstrated that the anomalously most probable importance for this Jürgen Sandkühler (Vienna, Austria) slow rate of desensitization removal function (evidence from KO animals roughly corresponds to the rate of inclusive, see also (Cockayne et al. Donald Simone (Minnesota, USA) unbinding of labelled ATP from the 2000)), it can be suggested that P2X3 William D Willis (Texas, USA) neuronal membrane (Pratt et al. 2005). receptors are most important in their

The authors demonstrated that to be heteromeric form with P2X2 receptors International Workshops 2007 desensitized, the channel has to be comprising a signaling unit which is Krakow, Poland (9-12 May) activated first. When activated, P2X3 “ever-ready” to generate long, weakly Lviv, Ukraine (18-23 September) receptor acquires a site which binds accommodating receptor potentials. ATP on a long-term basis. The affinity According to our experience, numerous Full details at http://www.physoc.org/international

Physiology News | No. 65 | Winter 2006 | www.physoc.org KIEV INTERNATIONAL WORKSHOP PN 33

Calcium disorders as a common denominator of different types of pain Within the last period, more and more attention has been paid to disorders of calcium homeostasis. These disorders at least accompany many types of neuropathic and inflammatory pain, and it cannot be ruled out that such disorders are one of the reasons for the development of many pain-accompanied pathologies

Long term chronic pain, mainly neurones independently of aetiological nociceptive stimuli. It has been shown inflammatory or neuropathic, afflicts diagnosis of pain. Molecular in recent works originating from about 25% of the general world mechanisms leading to changes in different laboratories that experimental population. More than 60% of people patterns of action potential generation chronic pain conditions (including aged 65 plus complain of daily pains. in primary neurones, modification of peripheral inflammation, nerve injury This degree of disability has a huge synaptic transmission between primary and diabetic neuropathy) do induce 2+ economic toll in terms of loss of central afferents and secondary dorsal significant changes in [Ca ]i regulation employment and disability payments, horn neurones and spinal neurone in DRG and DH neurones participating but quality of life is equally hyperexcitability acting in concert in transmission of nociceptive signals compromised. Pain is thus a major contribute to the generation of chronic Voitenko et al. 1999; Voitenko et al. medical issue; it is not simply a pain. In spite of a profound difference 2000; Huang et al. 2002; Kruglikov et sensation but an event that also triggers in patterns of changes or damage in al. 2004). aversive and threatening psychological primary and secondary nociceptive feelings. Patients in pain are likely to neurones the pain syndromes as Thus, impaired cytosolic free calcium become depressed and anxious, have observed in numerous studies regulation may be a basis for abnormal disturbed sleep patterns and generally conducted on different animal models nociceptive signalling during various have a poor quality of life. The are generally common. Thus, there is a types of diseases and pain syndromes. importance of understanding the ways reason to believe that multiple and Moreover, we have shown that these by which the central nervous system common mechanisms may be present in changes in the calcium regulation are can alter incoming signals that relate to chronic pain conditions of all similar in DRG and DH neurones pain processing stimulated an aetiologies. indicating that they might be general increasing number of investigations, for many types of neurones from many of which are concentrated on two Calcium is a universal second central and peripheral nervous systems. first elements in the pain pathway – messenger, and changes in the 2+ dorsal root ganglion (DRG) and dorsal intracellular cytosolic Ca Abnormalities in neuronal calcium 2+ horn (DH) neurones. concentration ([Ca ]i) triggers a wide homeostasis, which lead to impairment spectrum of cellular responses, of nociceptive neurotransmission, have Multiple, and possibly common, including a long-lasting modification of been detected in many pain models mechanisms appear to be involved in synaptic transmission and changes in such as diabetic and nerve-ligation­ the generation of chronic pain on the cellular excitability that may lead to induced neuropathies, carrageenan- and level of primary and secondary sensory changes in the transmission of CFA-induced inflammation, cancer

Figure 1. Nimodipine-induced restoration of calcium signaling and facilitation of diabetes­ induced sensory abnormalities. A and B, Representative examples of the effect of 2+ nimodipine treatment on amplitudes of [Ca ]i transients induced by application of caffeine (A) and ionomycin (B) in DRG neurons. The traces recorded from a neuron of control animal are shown in green, diabetic (red) and nimodipine­ treated diabetic (blue). Grey bars in B indicate application of Ca2+-free extracellular solution. C and D, Average values of amplitudes of caffeine- and 2+ ionomycin-induced [Ca ]i transients in DRG (C) and DH (D) neuronal preparations of control (green bars), diabetic (red bars) and nimodipine-treated diabetic (blue bars) rats. E, Hot Plate test: nimodipine treatment (blue bar) partially normalized thermal sensitivity in diabetic rats (red bar) as compared with control animals (green bar). Diagrams reflect a latency of nociceptive reaction at 48oC. * P < 0.05 vs. control, # P < 0.05 vs. diabetic animals. In all experiments n > 10.

Physiology News | No. 65 | Winter 2006 | www.physoc.org 34 PN KIEV INTERNATIONAL WORKSHOP pain, etc. In particular we have shown an increase of Са2+ entry to the cytosol via glutamate receptors in neurones of inflamed rats (Voitenko et al. 2004a); an increase of Ca2+ entry to the cytosol via voltage operated calcium channels in neurones of neuropathic rats (Voitenko et al. 2000); a decrease of calcium uptake and release by mitochondria (Svichar et al. 1998; Voitenko et al. 2004b) and a decrease in calcium mobilization from the different types of the endoplasmic reticulum (ER) calcium stores (Kruglikov et al. 2004; Voitenko et al. 2004b) in neurones of both pathologies. It is also important to emphasize that calcium abnormalities in these models are well correlated with the Figure 2. A simplified scheme of calcium theory of pain. An increase of Ca2+ entry into the cytoplasm together with development of many pain-related decrease of Ca2+ accumulation by the ER and mitochondria lead to calcium overloading. That, in turn, results in symptoms such as thermal and hyperexcitability of nociceptive DRG and DH neurons and changes of synaptic efficacy between them. All together it might produce central sensitization and pain. mechanical hyper- and hypoalgesia and changed pain sensitivity, indicating a nociceptive synaptic transmission and Kruglikov I, Gryshchenko O, Shutov L, Kostyuk E, Kostyuk P & possible relationship between them. Voitenko N (2004). Diabetes-induced abnormalities in ER calcium induction of pain. Therefore any mobilization in primary and secondary nociceptive neurons. Pflugers advances in the development of a Arch 448, 395-401. The other corroboration of the calcium unifying theory that may explain how theory of pain is the fact that chronic Shutov L, Kruglikov I, Gryshchenko O, Khomula E, Viatchenko- different diseases result in common Karpinski V, Belan P & Voitenko N (2006). The Effect of Nimodipine on treatment with calcium antagonists may pain abnormalities look very attractive. Calcium Homeostasis and Pain Sensitivity in Diabetic Rats. Cell Mol be effective in both the restoration of Neurobiol. (Epub ahead of print). calcium signalling in DRG and DH Based on many recent observations, Svichar N, Shishkin V, Kostyuk E & Voitenko N (1998). Changes in nociceptive neurones and the reduction mitochondrial Ca2+ homeostasis in primary sensory neurons of including those mentioned above, we diabetic mice. Neuroreport 9, 1121-1125. of diabetes-induced sensory venture to suggest the following abnormalities. We have shown recently simplified scheme that might be a basis Voitenko NV, Kostyuk EP, Kruglikov IA & Kostyuk PG (1999). Changes (Shutov et al. 2006) that treatment with in calcium signalling in dorsal horn neurons in rats with streptozotocin­ for this theory: increase of calcium induced diabetes. Neuroscience 94, 887-890. the calcium channel blocker nimodipine entry via voltage- or ligand-gated 2+ was able to restore impaired Ca calcium channels and/or decrease of Voitenko NV, Kruglikov IA, Kostyuk EP & Kostyuk PG (2000). Effect of release from the ER, induced by either 2+ streptozotocin-induced diabetes on the activity of calcium channels in accumulation of Ca by the ER and rat dorsal horn neurons. Neuroscience 95, 519-524. activation of ryanodine receptors or by mitochondria as a result of disease receptor-independent mechanism in development lead to temporal or Voitenko N, Gerber G, Youn, D, & Randic M (2004a). Peripheral both DRG and DH neurones (Fig. 1). inflamation-induced increase of AMPA-mediated currents and Ca2+ permanent calcium abnormalities. transients in the presence of cyclothiazide in the rat substantia The beneficiary effects of nimodipine That, in turn, results in hyper­ gelatinosa neurons. Cell Calcium 35, 461-469. 2+ treatment on [Ca ]i signalling were excitability of DRG/DH neurones paralleled with the reversal of diabetes­ Voitenko NV Potapenko I. & Shyshkin VO (2004b). Changes of and/or changes of synaptic efficacy intracellular calcium-regulating mechanisms of primary and secondary induced thermal hypoalgesia and between them. All together it might sensory neurones in periferal inflammation. Fiziol Zh 50, 33-41. normalization of the acute phase of the produce central sensitization and pain response to formalin injection. (Fig. 2). Nimodipine treatment was also able to shorten the duration of the tonic phase Acknowledgement of formalin response to a control value. Supported by JDRF grant #1-2004-30. It should also be noted that at the Nana Voitenko moment there is no single theory which Bogomoletz Institute of Physiology can explain the development of Kiev, Ukraine allodynia and hyper- and hypoalgesia during diseases of different aetiologies, References as well as no unified point of view Huang TJ, Sayers NM, Fernyhough P & Verkhratsky A (2002). concerning the influence of altered Diabetes-induced alterations in calcium homeostasis in sensory neurones of streptozotocin-diabetic rats are restricted to lumbar calcium homeostasis on neuronal ganglia and are prevented by neurotrophin-3. Diabetologia 45, 560­ Nana Voitenko at work in the Bogomoletz Institute of structures responsible for inpaired 570. Physiology.

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involving The Society, Editors and Blackwell Publishing, a new design was agreed and approved at the Board How to keep the loyalty of meeting (see example right). The Journal will continue to keep its by­ authors and readers volume colour scheme for the spine and this will form the background to the What is needed these days to keep a title and other front cover details, journal fresh and interesting to authors keeping them clearly readable and and readers? This question drove much distinct from the background cover of the discussion at the recent Journal image. of Physiology Editorial Board meeting in New York in September. At every The Journal is also introducing a new meeting the Board reviews standards mini-review article type aimed at and and Editors are encouraged to accept for younger scientists. The model for only papers that make an important step this is the successful J Neuroscience forward in their fields. In addition, for Journal Club feature and this title will a number of years the Board has be adopted for the J Physiol articles to commissioned timely Topical Reviews Senior) Editors. The review process acknowledge the inspiration for the in emerging new areas, sometimes was paper-based long after type setting initiative and inform potential authors associated with sponsored Symposia. and copy editing went digital. Postage about the purpose of the article. Journal More recently, recognising that bills were enormous – £25K by the Club articles will be short reviews of progress in science is the result of time that the online submission system recently published manuscripts and robust and frank debate, Editors have was introduced. Paper and associated authorship will be limited to graduate discussed introducing features that postage costs were only eliminated with students, post-doctoral fellows and involve an interactive element. Letters the introduction of the Bench>Press other training level scientists. It is to the Editor were introduced in 2004 online submission system in 2002 – envisaged that these reviews would to enhance interaction with readers. which strangely enough cost exactly the provide an appraisal/opinion of a amount saved in postage. The process manuscript published within the last It has been acknowledged that young of educating Editors in the use of the two issues of The Journal, and will scientists in particular, and especially online system has sometimes been provide young scientists with an young American scientists, need to be painful for both parties, but Ann has opportunity to hone their writing skills reminded that The Journal is a dynamic ensured that all but the most andintroduce them to The Journal of and forward-looking publication. electronically challenged Editors have Physiology early in their career. learned how to report online. Ann is One aspect of The Journal that can Other initiatives to attract readers and committed to making the process of deliver this message is the style of the enhance the appeal of The Journal will submitting a paper to The Journal of title. Earlier in the year The Journal’s be considered in the coming year. Physiology a positive experience. As Executive Committee agreed that the one Society Officer recently title style was beginning to look very commented, she makes being rejected different from those of current Milestones for Publications seem almost a pleasure. competitor journals and that the elongated serif font emphasised the Office staff In November, another member of the historical importance of The Journal On 1 September this year Ann Watson, Publications Office team, Linda rather than its current relevance. The well known to all Journal of Rimmer, will also reach her twentieth title was last redesigned in 1994 when Physiology Editors and referees, anniversary with The Journal and The The Journal changed from monthly completed 20 years on the staff of The Society. Linda began working for The hardback issues to fortnightly Journal’s peer review team. Over the Journal in November 1986, as a paperback issues. In the intervening years Ann has seen many changes to Publications Assistant in The Journal’s years The Journal has been the way papers are reviewed. She offices, which were then part of increasingly challenged to maintain the began work as a Distribution Assistant Cambridge University Press. The work loyalty of its authors and readers. With for Tim Rink, who was then one of the involved coordinating the flow of these thoughts in mind, the Executive Distributing Editors (DEs) based in the submitted manuscripts between DEs Committee decided to incorporate Cambridge Physiology Department. and The Journal office. Linda has since elements of The Society’s rebranding She worked for four Cambridge-based gone on to work with six Chairs and project (see p. 46) into a modernisation DEs before moving to the Publications one Editor-in-Chief. In 1988 she took of The Journal’s title and the company office at CUP in 1994, to form the on administration of the Editorial Board that has undertaken the project, 35, was nucleus, and eventual leader, of a team meetings and has organised around 40 commissioned to produce a new title. of Publications Administrators working Board meetings. In 1999 Journal of After a process of consultation remotely for the Distributing (now Physiology Symposia were introduced

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on development of antiarrhythmia therapies using implantable device approach.

New Editors His research laboratory employs and Three new editors have been appointed develops various optical imaging with an interest in the field of techniques to study action potential Computational Physiology generation, propagation and failure, including fast fluorescent imaging with Blackwell Publishing representative from Boston, voltage-sensitive dyes, optical Robert Harington with new Editor Daniela Pietrobon coherence tomography, quantitative from Italy (above); Editor Stefano Vicini (below) immunohistochemistry.

George Billman and these too have all been organised George received his Ph.D in Physiology by Linda (pictured below with Journal and Biophysics from the University of International Editor Steve Mifflin). The Kentucky, Lexington, Ky. After the success of these Symposia was recently completion of post-doctoral training Nic Smith acknowledged when The Society under the direction of H Lowell Stone Nic is a University Lecturer and Fellow agreed to increase the budget so that at the University of Oklahoma, he of St Johns College at the University of more can be organised. More recently, joined the Faculty of the Department of Oxford in Computational Biology. His Linda has become known to Society Physiology at The Ohio State research interests include the Members as the Executive Editor of University in Columbus, Ohio. He is development of integrated Physiology News. In this role she is currently, a Professor of Physiology and computational models which span Cell Biology there. His primarily cellular through to tissue scales. His research interest involves the work is this area has focused largely on investigation of the mechanisms the heart and specifically the coupling responsible for ventricular fibrillation. of coronary blood flow to excitation­ He currently is investigating the effects contraction under normal, acidotic and of endurance exercise training on ischemic conditions. cardiac autonomic regulation and the susceptibility to ventricular fibrillation induced by myocardial ischemia. November issue credited with raising the standard of the magazine while reducing its production costs. She has also been instrumental in reviving The Society’s Monographs Committee as the Biomedical Publications Committee, which is in the process of setting up a text book deal with Taylor and Francis, and producing many of The Society’s publications.

The contribution of both Ann and Linda Igor Efimov to The Journal and The Society over Igor is an Associate Professor of 20 years was marked by a presentation Biomedical Engineering, Cell Biology at the Editorial Board dinner. and Physiology, and Radiology at Washington University in Saint Louis. Carol Huxley His research is focused on the (photographs by Carol Huxley and BuckandTipp.com) mechanisms of cardiac arrhythmias and

Physiology News | No. 65 | Winter 2006 | www.physoc.org AFFILIATE NEWS PN 37

stereotypical image of scientists as well scientist. The meeting also incorporated Young scientists and the as ensure better science reporting. an insightful presentation given by Ted Griffiths (Director of the Biomedical media Selina Pearson Affiliate Member, University of Birmingham, UK Research Education Trust) on the need Like many scientists, I often get to deliver informed talks in schools on frustrated when I read or hear a science Sense about Science has recently animal research, and gave participants a story in the media. For a long time, bad published a media guide based on their feel of how little unbiased information science in the media has been workshops called Standing up for science on such matters is currently available in acceptable. Fortunately, however, which can be downloaded from the web site schools. The meeting, of course, things are improving. What can we do www.senseaboutscience.org/VoYS. involved more than just science, and to help improve the situation? taking advantage of vast nightlife available in London, the participants Scientists and the media are both at From molecules to enjoyed an evening of modern French fault, mostly due to the lack of food and drinks in a Spanish bar. The understanding that exists between the behaviour meeting concluded with prizes awarded two. This is where Sense about Science, The Young Physiologists’ Symposium to Clare Howarth (University College a charitable trust who promote the use took place alongside the main London) for the best talk and Nick of scientific evidence in public debates, Physiological Society Meeting in July Riddle (University of Cambridge) for come in. Through their VoYS (Voice of 2006 in sunny, bustling London. The the best poster presentation. Young Science) programme they run meeting, From molecules to behaviour, All in all the meeting provided a good VoYS media workshops for early career brought together young investigators opportunity for young investigators to scientists (post-doctoral fellows or from all over the UK. Presentations present their work and meet many equivalent in first job, and post­ covered diverse topics – from studies of people at similar stages of their graduate students) which aim to give ion channel structure and function scientific career in a friendly them an understanding of how the through to fMRI of human subjects atmosphere. We would like to thank media works. I attended their most and, of course, food, alcohol and sex The Physiological Society (especially recent workshop in June with other (α-MSH and appetite for sex in hungry Donna Brown) for giving us the early career scientists from the male rats from Celine Cacquineau of opportunity and the means to host such biomedical sciences. the University of Edinburgh and a meeting as well as everyone in the Evidence that ethanol equilibrates Physiology Department at UCL who The workshop consists of a series of rapidly across the C Elegans cuticle helped with the organization. Last but panel discussions. Our first panel from Philippa Mitchell of the not least, thank you to all the discussion was with a group of University of Southampton). The participants. We hope to see you scientists who had spoken to the media plenary talk, to an over-flowing lecture around! on controversial aspects of science. theatre, was delivered by Eve Marder They explained the problems they had (Brandeis University, Massachusetts). Izumi Fukunaga faced and how they had dealt with Together with Frances Edwards (UCL), Clare Howarth them. Eve formed the panel for judging the Rosemary Milton presentations and participated in many Ede Rancz We then spoke to a group of journalists. discussions on science and on life as a University College London, UK For most of us this was our first experience with the media, and they explained how they approach a story, what problems they encounter, and the pressures they are under. The final discussion covered the communication of (preferably published) research to the media, the value of media training, and the importance of communicating accurate information to the public.

It was a highly informative and enjoyable workshop, directed at younger scientists, who tend to be in the lab more and are usually more nervous about talking to the media. We were encouraged to get more involved in scientific debates which we all agreed would help challenge the Shmma Quraishe (left) and Sarah Luedtke from Southampton University who participated in the YPS.

Physiology News | No. 65 | Winter 2006 | www.physoc.org 38 PN MEMORABLE MEMBERS

matter of disappointment rather than David Henry Smyth celebration – ‘it’s like getting a CBE Ian Roddie looks back at the when expecting a knighthood!’ distinguished physiological career of ‘an adult white rat University administration did not attract him but he was a natural choice as enterocytologist’ chairman for any committee whose length was always shortened by his David Smyth was born in Lisburn in brief and pungent summaries of Northern Ireland in 1908 where his evidence. His perceptiveness, good father was a primary school judgment and, not least, his ability to headmaster. After reading medicine in express himself clearly and directly Belfast and a postgraduate studentship made him one of the wisest counsellors with Herman Rein in Gottingen he and led to a four year stint as Pro-Vice proceeded to a distinguished career in Chancellor. Though bespectacled and academic physiology. relatively short in stature with a voice It began with a lectureship at UCL with David Smyth – a hard worker, original thinker, witty characterised by a stammer and Charles Lovatt Evans. When war and humorous, modest, good-natured and passionate pronounced Ulster accent, he was much started in 1939, he was evacuated with about the Keeshond (pictured below). in demand as a speaker. At Society part of the UCL medical school to dinners his wit, modesty, commonsense the widest possible scale. The person and humour were such that his Sheffield. Here he worked with the who plays one game intensively and future Nobel Laureate, Hans Krebs, audience gladly forgave any problems sticks to this may learn more of in delivery. who summarised his character with gamesmanship than the one who remarkable astuteness: ‘We all learnt dabbles in many. The gardener who His skills and attributes were very quickly to appreciate his specializes in chrysanthemums or recognised widely by The Society. He exceptional qualities. Not only was he dahlias or roses will get greener fingers was onetime Chairman of the Editorial a very hard worker, he was also an than the person who keeps the whole Board of The Journal of Physiology original thinker, always helpful, very garden tidy. The best musicians have (1966-68), Foreign Secretary of the witty and humorous in his comments on usually started and many continue to Society (1972-79), Chairman of British day-to-day occurrences. Modest in his concentrate on one instrument. And if I National Committee for Physiological bearing and good-natured, he never may give another example from one of Sciences and the British delegate on the complained. In spite of being busy he my own hobbies – pedigree dogs – the Council of the IUPS. was never in a hurry or “flap” and person who knows most about dogs in always had time for kindness.’ general starts by concentrating After retirement he continued work on intensively on one breed. But perhaps I intestinal absorption and busied himself When the UCL School moved to am only making excuses for the most with writing. He wrote eight ‘Personal Leatherhead in 1940, with FR Winton selfish and yet perhaps the most View’ articles for the British Medical as Dean, David became Sub-Dean. In fundamental of all urges – the urge to Journal that became famous for their 1946 he moved to the Chair of do well with all your heart and soul wit and insight. He wrote a book, Physiology at Sheffield where he what you really love doing.’ Alternatives to animal experiments, in remained until retirement in 1973. For 1978, and succeeded Sir William Paton his work on intestinal absorption he David was passionate about rearing as Chairman of the RDS. He was was elected FRS in 1967. pedigree dogs, particularly the Dutch involved in drafting Lord Halsbury’s Barge Dog, the Keeshond. The Smyth Laboratory Animals Protection Bill and In all matters he set himself name is still mentioned with near chaired the committee of the University exceptionally high standards. Central reverence whenever specialists talk of Federation for Animal Welfare. to his philosophy was the need to Keeshonds. On being congratulated concentrate in depth in one field rather when one of his dogs won second place In 1979 colonic cancer was diagnosed. than range over many. at Crufts, he demurred saying it was a Though the surgery was deemed successful his condition deteriorated Unapologetically, he described himself and he failed to recover. His many as ‘an adult white rat enterocytologist’ friends attended the funeral at Eyam and defended his policy thus, ‘I have Parish Church near his Derbyshire tried to defend the position of the home. extreme specialist through the paradox that only he who looks deeply into one Ian C Roddie small problem really appreciates some Honorary Member, London, UK of the universal generalisations that lie below. I believe this happens in life on Next time, Liam Burke remembers his time in Bernard Katz’s laboratory at UCL in the 1970s.

Physiology News | No. 65 | Winter 2006 | www.physoc.org THE PHYSIOLOGICAL SOCIETY LECTURES PN 39

The Bayliss-Starling Prize Lecture

agog with the exciting experience.’ So Evans became Sharpey Scholar at £150 per annum. That must have been a substantial sum for those times. So began a new life at UCL.

Bayliss was born in 1860, and Starling in 1866, so 1963 was the mid-point between their centenaries. Starling died at 61 and Bayliss at 64.

William Bayliss ‘Bayliss was well-to-do, but of simple Ernest Starling personal tastes, and of the most gentle, The first lecture in this series was given retiring and kindly disposition. He had Starling who was then at Guys. They by Sir Charles Lovatt Evans who had a large house in a 4 acre garden ‘were complementary. Bayliss was been Jodrell professor and head of the adjoining Golders Hill Park’ where patient, over-modest and most genial. Department at UCL from 1926 to 1949. there was tea and tennis on Saturdays. He radiated happiness and his talk was Entitled Reminiscences of Bayliss and He was married to Starling’s sister. In a spiced with laughter. … his reading was Starling it was a beautiful account of a shrubbery between the two tennis vast and omnivorous.’ With him it was world long lost, even in 1963. The courts was a small summer house not master and pupil, but two lecture was published by The where … one might find Bayliss colleagues comparing notes. In 1922 he Physiological Society in 1964 and I surrounded by piles of penny exercise received an instruction ‘to appear at treasure my copy. I believe it was sent books, suggesting a marking of Buckingham Palace to receive the to all Members of The Society. The homework. This was in fact the accolade of the Order of Knighthood; quotations below are from this preparation of the Principles of he replied that as the date coincided published lecture. physiology … Close by was a with that of a Meeting of The laboratory he had built years before, Physiological Society, he would be I first met Sir Charles in 1958 when I and in which he afterwards made the unable to attend’. His priorities surely was drafted into the RAMC and sent as photographs for the illustration of his were right. a physiologist to Porton Down. He had book…’. This book, in the early retired from the Jodrell Chair at UCL editions, is a marvellous monument to In 1903 he was involved with the anti­ and was head of the physiology section. his erudition. He had studied science at vivisectionists in what became known He came in to the department some 3 UCL and then as a medical student as the ‘Brown Dog case’. He was days each week and invariably had tea failed anatomy. At the oral examination accused of performing an experiment or coffee and a chat in the lab in which the examiner said : ‘your written on a dog without anaesthetic. He won I worked. So there were many stories answer to the question on the cords of an action for libel, with £2000 about UCL, many of them scandalous. the brachial plexus was extraordinary – damages, against Stephen Coleridge, The obituary by I de Burgh Daly and R wherever did you get it from?’ To which sum he gave to the college. A Gregory is a charming and which Bayliss replied ‘Well I never Bayliss was exposed to ‘the usual sympathetic account of his life could memorize that sort of thing, so I accompaniment of publicity, prejudiced (Biographical memoirs of Fellows of put it the way I thought it ought to be.’ press comments, and shoals of the Royal Society 1970, 16, 233–252). By 1890 he began to collaborate with anonymous letters, abusive, threatening,

Lovatt Evans had worked with Bayliss and Starling from November 1910 when he had been an external candidate for the London University BSc in physiology and the practical examination was at UCL. Francis Gotch was the external examiner. Gotch asked Evans to stay behind as Starling wished to talk with him. Thus he met the staff and ‘I was quite overcome by the natural friendliness of all these A rubbing of the plaque erected at UCL to commemorate the Institute of Physiology (courtesy of the Physiology well-known people, and returned home Department, UCL)

Physiology News | No. 65 | Winter 2006 | www.physoc.org 40 PN THE PHYSIOLOGICAL SOCIETY LECTURES obscene or comical, the case must have been very distasteful to him at the time …’. The best account that I know of this case was written by Leonard Bayliss, son of W M Bayliss, The ‘Brown Dog’ Affair printed in a UCL Physiology Department magazine entitled POTENTIAL (Spring 1957, No 2, pp 11-22, UCL, Price 1/-). This piece is hard to find.

A bronze statue of a dog was erected in Battersea by the anti-vivisectionists to commemorate a dog. The statue was subsequently removed after attacks by medical students. The council was sued but the subsequent action went in the favour of Bayliss given the libellous nature of the inscription. The statue was erected in Latchmere Gardens and the cast iron fence around the area carried a A reconstruction of William Bayliss’ sign in the 1980s saying, apparently lecture used in the famous 1903 libel without irony, No dogs allowed. case (above, left); the ‘Brown Dog’ Photographs of the re-staged statue in Latchmere Gardens, experiment, of the statue of the dog, Battersea, erected by the anti­ vivisectionists (above, right); the site and of the location can be seen in The where the statue once stood (left). world of UCL by Harte and North (revised edition 1991, p. 127). was 19 years old at the time. From both that the Physiology Laboratory was in Charles Darwin (an original Honorary the North wing of the College above In contrast with Bayliss, Starling was Member) and T H Huxley (a founder the Slade School. It is commemorated ‘brisk, ambitious, a bit quixotic, Member) ‘he drew the wide biological at the top of the stairs in the front east serious, generous and highly strung … vision which characterised his writings, corner of the Slade School. From the he enjoyed the limelight, and the use of and from the second a keen interest in basement ascend nearly 75 steps to the power, for good ends, but was too education and the desire to smite all second floor. There is a grey stone forthright and impatient to be humbugs – however big.’ plaque with engraved inscription tinted diplomatic, though he realized that gold on the wall to the rear. The power lay that way … he enjoyed He was elected to The Physiological inscription reads ‘This was the company, good wine and food, dancing Society ‘on the same day as Bayliss, at department of Physiology 1881-1909. and tennis, and was always keen on the Annual Meeting at University Here worked Sydney Ringer J Burdon helping young people.’ College on 15 February 1890, Sidney Sanderson Edward Sharpey Schafer Ringer in the chair’. John Rose Bradford William Bayliss ‘At the Physiological Congress in 1926, Ernest Henry Starling’. G H Hardy he (Starling) said that the British Starling, aged 33, went to University would hae regarded this as forming the excelled in science because they were College as Jodrell Professor in 1899, foundation of a very good first 11. devoted to sports, and research was the the year of his election to the Royal greatest game of all. At which Gley, a Society. Starling was dissatisfied with this small man with the voice of a bull, in accommodation and there came to be an impassioned oration described And so the present Institute of built a large new department to be a France as “le berceau de la Physiology came to be built. Before nucleus of the new Faculty of Medical physiologie”, and Pavlov spoke in German of his own invention.’

Starling’s background was in medicine The 2006 Bayliss-Starling Prize Lecture, with the fundamental science added Annexin 1, glucocorticoids, stress and ‘largely through his association with inflammation, will be delivered by Rod Flower, Bayliss, but the needs of medicine FRS (William Harvey Research Institute, UK) really formed the foundation of most of (pictured right) at The Physiological Society his work’. He was much influenced by Focused Meeting at the University of Bristol on Wooldridge at Guy’s who advised him Monday 4 December 2006. to work with Kuhne at Heidelberg. He

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Sciences. It was located on the land even their first publication, on the investigations of enzyme action, previously forming the playground of electromotive phenomena of the heart adsorption, and surface action. Starling the University College School for Boys beat, was of pioneer quality. introduced the heart – lung preparation which moved to Hampstead. The Fundamental, if less dramatic, and and enunciated the ‘law of the heart’. building was completed within 1 year bearing valuable, if late, fruit, were Starling published The Principles of of going to the architect and with all Starling’s own work on lymph Human Physiology and Bayliss equipment and fitting out cost formation and on renal excretion, and Principles of General Physiology. ~£20,000. Large gilt letters on the the importance of Bayliss’s ‘Both books served to set the subject front proclaim Institute of Physiology investigations on membrane potentials into its present mould.’ in an art nouveau style to be seen today. needs no stressing in these days.’ This was a tribute to German ‘Bayliss, the serene and devoted physiology institutes and ‘much The discovery of secretin and the scholar, the true natural philosopher, disliked by the then Provost, who coining of the name hormone, came Starling the man of action, with wide thought he scented secession’. The about in an environment where the sweeping views; neither of them were building was very advanced for the notion of chemical transmission was subjected to the isolation of the time and in 1909 Starling wrote a discussed. Oliver and Sharpey had specialist. It might be said that Bayliss Description of the new physiology published their work on the adrenal and Starling flourished at a gestatory institute. gland, and Eliot was contemplating period in the history of physiology, a ideas about the autonomic system. The period in which, out of the fullness of Lovatt Evans summarised the joint time was right for their beautiful time, the emergence of modern work of Bayliss and Starling thus: ‘The experiment and brilliant insight which physiology was due.’ fruits of their labours have by now been opened up a whole new field of homogenized into current knowledge, biological science, not only in medicine These lectures justifiably continue to but the source of their enrichment and physiology. honour them. should not be forgotten. The discovery of secretin was no doubt their most There was also Bayliss’s work on the Tim Biscoe spectacular and fertile contribution, but vasomotor system and his Honorary Member, London, UK

A life of Ernest Starling the early history of physiological research at Starling also emerges as a believer in science Guy’s Hospital and University College, the as something that transcended national By John Henderson institutions where Starling spent most of his boundaries. Apart from the British physiologists 2005, Oxford University working life. The book follows Starling through who appear in the book, international figures of Press. £35.99 (hardback) all the phases of his career, including his war the time like Pavlov and Gleb Anrep also work in poison gas defence and his key appear, testifying to the internationalism of ISBN 0 19 517780 0 influence on medical education in the London physiological research both before and after the colleges. Great War. Like many of his contemporaries, When reviewing a biographical memoir, one is Starling had trained in the laboratories of the inevitably faced with the question of how much Starling comes across in this book as a great German physiologists of the late 1800s biographical information to include. In the case surprisingly modern figure in some ways, and and was by his early 20s a fluent German of John Henderson’s entertaining life of Ernest often notably out of step with the more genteel speaker. He thus felt deeply betrayed when Starling (1866-1927), published by OUP as part tenor of the times. Tim Biscoe quotes Lovatt Germany went to war in 1914, penning wartime of a series sponsored by the American Evans, speaking in 1963 many years after letters with scathing references to ‘the Hun’ and Physiological Society, Tim Biscoe’s extended Starling’s death, describing Starling as ‘highly advocating the use of Mustard Gas as an introduction to the Bayliss-Starling Prize strung’ and ’forthright and impatient’. The offensive weapon. But after the war he seems Lectures above saves me the trouble, and can book’s picture of Starling bears this out, to have felt that what was important was for be consulted for details. suggesting a man given to saying exactly what scientists to get back to doing science, and he he thought, unvarnished, and without much welcomed the German physiologists back into Starling is a fascinating figure. Although there is regard for how it would be received. Through the fold and continued to publish in German plenty of competition, he must be a leading his outspokenness Starling seems to have had journals. This forgiveness was not universal in candidate for the title of ‘most important the knack of offending various establishments, science at the time, and may even have cost physiologist never to have won the Nobel medical, military and political, which almost Starling the Nobel. But it gives a true flavour of Prize’. He was also never knighted, despite his certainly cost him a knighthood. Even when the man and his principled, if rather black and pre-eminence and unlike many of his being complimentary Starling does not seem to white, views. contemporaries. To learn why, read this book. have been the most tactful of men. The book describes how he recommended A V Hill, who To sum up, John Henderson has given us a Henderson’s book is enjoyable both for its he greatly respected, as his successor as fascinating portrait of a great scientist, and a glimpse into the past and for the contemporary Jodrell Professor of Physiology at UCL, but told man of tremendous conviction, though poor parallels it tempts one to draw. Henderson the Dean that Hill [was] ‘not a physiologist … of political instincts. Readers will enjoy both the recounts Starling’s scientific achievements, and course’ a remark clearly capable of being history and the contemporary parallels, and the his collaborations and interactions with other misinterpreted (the point was actually that Hill book deserves a wide circulation. scientific eminences like Bayliss, Charles was something different, namely a Lovatt Evans and (later) A V Hill, and describes biophysicist). Austin Elliott

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author fee for its journals to $2,500 and is was not greatly missed and was Parliamentary and Scientific still not able to balance the books, in spite presumably being accessed successfully by Committee of $13 million in donations. He also pointed those who needed it. out that variants of OA already exist: delayed OA, as exemplified by the society The wide variety of views on the question of journals, whereby all content is free after a open access was made clear during the period, e.g. 12 months as is the case for discussion after the presentations. If a The Journal of Physiology and Experimental researcher works in a well-funded institution Physiology; free access to developing in a well-funded discipline, OA does not countries through special agreements made present too many problems. For many it is with the publisher. He urged that proper not a realistic possibility; if the funding to research like the study proposed by RCUK pay author fees is not available, getting should be conducted before forcing a published will be a major challenge. change in the publishing model and pointed Similarly, learned societies that rely on out that high quality journals would probably subscription income to fund their survive a subscription period of 6 months programmes will be challenged to replace but lower quality journals would not and that income. On the plus side for OA, it was many society journals would be at risk. And noted that it allowed publication of negative he finally made the point that in the UK we results, which will restore the balance to have a very successful knowledge-based clinical studies. And barriers to readership economy and that the industry behind this will be lifted. It was pointed out that should be supported. researchers doing interdisciplinary work The July meeting of the Parliamentary and cannot access the information they need Scientific Committee asked the question ‘is Ian Rowlands and David Nicholas (Centre because their libraries cannot afford the full open access the future for scientific for Information, Behaviour and the range of journals. Mark Walport mentioned publishing?’. Evaluation so Research (CIBER) at UCL) that there are other ways of making content reported on their surveys of author attitudes accessible that can co-exist with the The first of four experts presenting to the to OA and existing OA models. A recent subscription model. Some journals are meeting, Mark Walport (Director of The CIBER author survey revealed that authors experimenting with very cheap pay per view Wellcome Trust), claimed that open acccess view OA differently depending on whether access models and societies might look at (OA) has arrived and is here to stay. they are considering their own science or this as an alternative way to raise funds Journals and learned societies must adapt. science in general . 74% agreed that high from their publishing ventures (although He stressed that the publication is an prices make access to journals difficult, but information from a recent HighWire Meeting intrinsic part of the research process and only 25% wanted to publish in OA journals; suggests that these experiments are not should be funded in grants and that it would 48% agreed that too much science is raising much income). On the other hand, on average constitute 1% of a research published, but only 13% agreed that they there are concerns about the model. Open grant. He also pointed out that submissions publish too much. The survey found little access proponents assert that peer review to the Oxford University Press experimental interest in, or understanding of, OA among will not be changed but lesser journals OA journal Nucleic Acids Research are authors. Attitudes to OA were heavily whose income depends on the number of holding up; authors are finding the money to dependent on discipline, age, funding and articles published may be tempted to pay the OA fee. geographical location. ‘Mainstream’ publish more. The barrier to readership will scientists were generally Anglo-US based, be replaced by a barrier to authorship; Robert Campbell (President of Blackwell ignorant, hostile and averse to change. ‘OA either way subsidies are required to ensure Publishing), an advocate of gradual change enthusiasts’ were largely young Asian, that the research output is published and is in the publishing models, made the points African or Eastern European clinicians, available to all who require it. Giovanni that there are serious concerns about the mathematicians, computer scientists or Mann spoke to the meeting about The OA author pays model. (1) There is the engineers. From the authors’ point of view a Physiological Society’s role in training potential for a reduction in standards. single model does not suit everyone. CIBER young physiologists and how this would not Publication costs money and corners may also reported on their study of downloads be sustainable if the subscription income have to be cut. Smaller journals will be from the OA journal Nucleic Acids Research from the journals was lost. Mark Walport tempted to publish more if article fees are both before and after the journal went fully responded that The Society should apply to their only source of income. (2) The barriers open access. The journal site was opened The Wellcome Trust for a training grant. Of to readers may be lifted, but there will be up to Google in 2003 and this had an course, the present costs of Society barriers to authors who cannot afford to pay immediate effect on downloads from the programmes for training young scientists far the fees. Access to the top journals will be site, which increased 143%. The exceed the amount The Wellcome Trust are confined to Northern hemisphere authors. subsequent increase in downloads when likely to grant. I believe Giovanni is framing (3) There is no evidence yet that OA the journal went fully open access in 2005 his reply. publishing is viable. The Public Library of was 8-10%. This suggests that the content Science (PLOS) has recently raised the that was previously not available via Google Carol Huxley

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threats to R&D. Dominance of Eastern widely applied in China, India and the Parliamentary Links Day low wage economies could undermine USA, but with almost no uptake in The Royal Society of Chemistry’s manufacturing industry in the West. Europe through inadequate addressing (RSC) Parliamentary Links Day on Politicians have sometimes looked to of consumer concerns and political science and globalisation held on 27 the service sector to plug economic pressures. His conclusion was that the June was an impressive event. Chaired gaps left by the decline of globalisation of science would continue by Brian Iddon MP, it was very well manufacturing, but service industries in spite of Luddites in some regions, attended by Parliamentarians and cannot be expected to invest in R&D in and that what had proved to be true for representatives of the scientific the same way as manufacturing. Over­ GM crops would also apply to other community. Our new Chief Executive regulation in Europe and the USA, for potentially controversial technologies Mike Collis sat with Sir Geoffrey Howe example REACH, might also drive such as stem cells. New technologies (now Baron Howe of Aberavon), and companies eastwards, places like have to be effectively managed rather the whole event was certainly an Singapore which are now investing than resisted. insight into what can be achieved by a heavily in R&D and infrastructure will learned society with enough clout – be only too happy to attract them. Dame Julia Higgins (Foreign Secretary perhaps something similar might be Companies like Government support, of the Royal Society, RS) stated that the considered under the umbrella of the access to an educated workforce and RS very much views itself as an BSF in future years. sensible tax regimes. It is vital that the international organisation. It actively UK provides this sort of environment to recruits foreign members, and provides The opening address was from encourage inward investment. funding to stimulate international Margaret Beckett MP (Secretary of collaborations including engaging State for Foreign and Commonwealth Robert Kirby-Harris (Chief Executive academics worldwide on major policy Affairs). She commended the occasion of the Institute of Physics, IOP) said issues. The RS spends about £7 million for being the most important scientific that physics is a major force for per annum on international interactions, event in the Parliamentary calendar. development, e.g. through education including a programme of international She felt that Britain’s standing abroad work in the developing world. It is a lectures, an MEP Pairing Scheme for was very much linked to our status as a core underpinning discipline for young scientists (based on the very scientific nation, but the global industry and medicine, for example successful UK MP Pairing Scheme) scientific landscape is shifting with the magnetic resonance imaging and which aims to increase awareness of emergence of India and China as being materials for transplants. The UK leads science in the European Parliament and possibly the next knowledge the world in laser physics, non-linear an understanding of Brussels amongst superpowers. China has trebled its optics and photonics. The IOP has scientists. The RS has been very active spending on R&D in the last few years made a serious investment in its in tackling development issues and India now produces more science international activities. They have including input to the G8 2005 joint graduates than the whole of Europe. offices in China, Japan and Russia. statements on Science and Technology This could be a positive development Their education projects include for African development, and has been for Britain if we seek to fully engage funding mobile labs in a lorry which very vocal to the Department for with them and build new tours Africa introducing science to International Development (DFID) on collaborations. Science thrives on young people, setting up training the role of science in eliminating diversity. The Foreign and programmes in entrepreneurship for poverty in Africa and elsewhere. Africa Commonwealth Office (FCO) Science developing countries and supporting is a priority for the RS at the moment. and Innovation network of attachés relevant African learned societies to They are strengthening the role of posted around the world is poised to create a pan-African network. African science academies to provide help us build these collaborations. advice to their Governments in Alan Malcolm (Institute of Biology) partnership with the Network of Simon Campbell (President of the presented a case history of a very African Science Academies, are RSC) stressed that the RSC was not controversial area of science and launching a new grant scheme for UK centric in its outlook and wants to globalisation – GM crops. He made the partnerships between UK and African be the most effective international point that this could be viewed as both scientists to promote capacity building organisation for advanced chemical a failure and a success. A decade ago, in the developing world, and have science. Various initiatives help them the technology had hardly impinged on funded several African academies to do this. They publish about 45 leading the international scene, now over one become members of the International textbooks every year, hold international million square kilometres are under Council of Scientific Unions (ICSU). joint meetings with the help of the GM cultivation. Momentum in this area FCO, have made their journals free to is considerable – more than 50% of Peter Saraga (Royal Academy of all African scientists, produce education soya produced worldwide is now GM Engineering) gave a personal materials for the developing world and herbicide tolerant, and about 25% of perspective from his experience of have an initiative called Chemistry Aid. maize is GM insect resistant. The working globally for Phillips. Industry He highlighted some of the potential technologies are very popular and needs to combine the best of global

Physiology News | No. 65 | Winter 2006 | www.physoc.org 44 PN PARLIAMENTARY LINKS DAY science and technology to create through uncertainty, and politicians He noted that the Government’s globally marketable products. Global need to be engaged in global science Foresight processes had changed to get businesses looking to access the best that addresses global concerns such as scientists and social scientists to work talents are looking towards Asia, for energy supplies and climate change. He together to tackle major issues such as example Phillips is locating more of its noted that there is still a problem of flooding and coastal defence. Other R&D in China, but they are still scepticism and ignorance about science Foresight exercises that he mentioned investing in good R&D in the UK. amongst some parts of the general included brain science, addiction and Successful knowledge transfer is based public, and gave perceptions of animal drugs, the determination and on user needs and he noted the research as being a particular issue. He identification of infectious diseases and emergence of a new trend in concluded that MPs have a key role to tackling obesity. international collaboration towards play in maintaining public confidence open innovation, with more emphasis in science. The session was wrapped up with a few on sharing knowledge as a route to complimentary remarks by Mark Lancaster MP, the son of the famous success rather than the more secretive Sir David King (Government Chief intellectual property route. firework maker the Reverend Scientific Adviser) ended the session by Lancaster, who was apparently in the highlighting that science and innovation Zhiyong Jin (First Secretary for Science audience. Alas, no firework displays was expected to play a key role in the were available on the day but they and Technology at the Chinese wealth creation agenda supporting vital Embassy in London) talked about consoled us with a good buffet lunch. I services such as pensions, education was left with much food for thought on Chinese Government emphasis on and health. The success of S&T has international co-operation in S&T ways of developing our Society’s been amply demonstrated through international activities and input to development. The Chinese economy is increased life span and an explosive booming and is becoming very Government policy. Amongst other growth in world population. How we things that I will be investigating in the dependent on S&T. Chinese S&T manage this in a limited resource development and reformation of the forthcoming months will be how we environment is the big challenge for can get our scientists involved in S&T system is directed by three main 21st century technology in water and strategies, in human resources, patents Government Foresight exercises and energy, health, medicine and other key other policy development activities. and standards. They are looking to areas. This is a big opportunity, and the strengthen audit and evaluation of Members with any ideas about this are Government is looking to support the more than welcome to contact me. research, develop new science policies, UK as the partner of choice for laws, regulations and public science international scientific collaborations. Liz Bell engagement. They are promoting knowledge transfer by new S&T intermediaries and incubators and high Introducing Affiliate Staff update tech industrial development zones. The Representative Lisa Mullen David Bennett (Events Administrator, Chinese are now very keen to support International) and Heidi Adnum international scientific collaboration (Events Administrator, UK) (below) and have signed inter-governmental have recently joined The Society’s S&T agreements with 96 countries, Events Team. Elfa Wilmot has been including the UK. There is a biennial made a permanent member of staff China-UK S&T Joint Commission (responsible for human resources, the Meeting which promotes collaborations Benevolent Fund and travel grants), and seeks to remove obstacles. His and Gabina Alfonso has been presentation was followed by Jason Lisa Mullen (above) became an appointed as full-time Finance Hahn (Science and Technology Affiliate Representative on The Administrator. Counsellor of the United States Society’s Council in May and will Heidi can be contacted on 020 7269 5715 Embassy), who noted that international serve a 1 year term. Originally a ([email protected]), David on 020 7269 science promotes the values of free secondary school teacher, Lisa 5712 ( [email protected]), Elfa on 020 societies and respect for diverse views, undertook an MSc in physiology and 7269 5713 ([email protected]) and and is essential in tackling issues of then went on to complete a PhD, Gabina on 020 7269 5710 global concern such as climate change graduating from Birkbeck, University ([email protected]). and epidemics. of London in 2004. Lisa is currently a post-doc at University College Phil Willis MP (Chair of the House of London. She sits on Council, Commons Select Committee on representing the views of Affiliate Science and Technology) commented Members and has yet to attend her that international co-operation in S&T first committee meetings this autumn. is now of vital interest to Parliament. She can be contacted by email on Science is about picking a route [email protected].

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Governance for beginners – The Society is eligible to stand. Council The 23 Members are elected to serve for a Council Members/Trustees the structure of The Society period of 4 years. The current size of the Council is 23, but is being reduced Jonathan Ashmore, Doug Corfield, The organisational structure of The to 20 as approved at the AGM in 2005. David Eisner (International Society’s governance can be a mystery Meetings of the Council are chaired by Secretary)*, Valerie Gladwell, Paul to Members. This article aims to the President. Greenhaff, Sarah Hall, John explain in simple terms how The Hanrahan, Patrick Harrison, Anne Society is run and how the decision In addition to Trustees, there are other King, Prem Kumar (Meetings making process works. invited attendees at Council meetings. Secretary)*, William Large (Editor-in- They contribute to discussions, but do Chief, The Journal of Physiology)*, Council of Members/Trustees not vote. These are currently: At the heart of the organisational Graham McGeown (Treasurer)*, Ian structure of The Physiological Society • two Affiliate representatives, McGrath (Chairman of the Executive is the Council. All major business of nominated and elected by Affiliates Committee)*, Stafford Lightman, The Society is directed by the Council, • Chair of the Editorial Board of Clive Orchard (Vice-Chairman of the Experimental Physiology which currently meets 3 times a year. Executive Committee)*, Ole Petersen • Editor of Physiology News (President)*, Christof Schwiening, Council Members are Trustees and, as • Chairman of the Heads of Sergey Smirnov, David Sugden, The Society is a charitable company Department Committee registered with Companies House, also Alexei Tepikin, Keith Thornbury, Administrative support is provided to Teresa Tiffert and David Wyllie. Directors of the company. Trustees are the Council and its Committees by The legally responsible for the overall Society’s staff based in the London and * members of the Executive Committee management, finances, administration, Cambridge offices. policy and strategic planning of The Physiology. The normal term of office Society, ensuring that the charitable Executive Committee on the Executive Committee is 4 years. objectives for which it has been set up The Council delegates the day-to-day are met. management of The Society to the Committees and Working Executive Committee, which meets Groups The Council consists of: frequently throughout the year. The Council also has Committees and Members of the Executive Committee Working Groups to which it delegates • a President, elected by the Council powers, and these groups report back to • one Member elected by the Editorial are drawn from, and elected by, the Council. the Council. Membership of these Board of The Journal of Physiology Committees and Working Groups is (normally the Editor-in-Chief); and There are seven members of the drawn from the Council, but can also • Members nominated by the Executive Committee: the President of include other invited members. The membership or by the Council and The Society, a Chairman, a Vice- Society’s Committee structure is elected by the membership at the Chairman, a Treasurer, a Meetings currently under review by the Annual General Meeting. Secretary, an International Secretary, Executive Committee. Elections to the Council take place each and the Member elected by the year at the AGM, and any member of Editorial Board of The Journal of Audit and Risk Committee The Audit and Risk Committee was set up to meet the requirements of good practice suggested by the Charity Commission. It reviews the effectiveness of internal financial controls and risk management systems, the effectiveness of our internal audit functions and the clarity and completeness of disclosures in the financial statements. The Committee consists of non-Executive Council members as well as invited external members. Simon Kellas Committees and Office Manager

For more information visit The Society’s web site at Current Committee structure http://www.physoc.org

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Society rebranding

As you can see from the images pictured (right) The Physiological Society has been represented by what can only be called a mixed bag of brands (used in the loosest sense!) over the years. Whilst some may argue brand doesn’t really matter to a society like ours and shudder at the mere thought of ‘re­ branding’, a small group of staff and Members thought differently, and sat down to review all Society publications – literature, stationery, and promotional materials. It should be possible to tell at a glance that something comes from The Society. Think BHF, Wellcome Trust … Our part of the biology curriculum) and physiology conclusion was simple – there was just no departments are rapidly being swallowed by consistency: logo, typeface, colour, imagery … Perception of The Society the trendier ‘biomedical sciences’ or such-like. the list goes on. Role • Support our Members The concept of branding was presented to the We agreed that we should contact • Support scientists Executive Committee and it was agreed that organisations that had themselves recently re­ • Promote physiology to schools and students the recommended agencies should be invited • Support and encourage physiology branded, and, not surprisingly, we found that to tender. Surprisingly impressed by the • Influence government and opinion formers their new looks were courtesy of several highly presentations, Exec invited the preferred • Promote the discipline and explain its importance recommended communications agencies. Initial agency, 35 Communications, to present to Vision consultations with three of these agencies Council who, after lengthy discussion, gave the • To be financially sound concluded what we already knew – that we • Provide quality and value to Members project the green light. (I should perhaps point were all over the place when it came to • Ensure subject remains relevant and recognised out that this is a simplified version of events branding and to increase the presence of The • To be more a voice of influence which in fact took 6 months to conclude.) Society we needed to do something about it. • Professionally led, strong Society • Increase public understanding of science Over the next 6 months the brand was Perception of identity Branding isn’t just a logo, something we could developed. The first step was to research what • Everything is inconsistent draw on a beer mat at the pub; it covers all The Society represented so 35 interviewed • Not attractive to a younger audience aspects of visual communication and ensures numerous stakeholders over several weeks. • What identity – we haven’t got one! that our presence is clear and easy to • Our identity has no lasting recognition The findings which were presented to Exec are understand no matter where or how it’s being • The old dog must go – irrelevant, too lengthy to summarised in the table (right). used. A brand also needs to be easy to apply explain with clear guidelines for all users, which in the Perception of current Happy they had an accurate view of The long-term saves time and money. communications Society’s role and core values, 35 developed • Needs to be more engaging numerous different concepts which they • Quality is varied and there is no system There are many reasons why we want to presented to a group consisting of Exec and • Everything we produce is different increase the presence and recognition of The other interested parties. The group was very • Communications are generally patchy and Physiological Society, all of which will benefit inconsistent impressed with many of their ideas which were our current membership, but one of the primary • Web site needs to draw in the younger members of separated under four headings (exploration, reasons is to ensure physiology is a recognised the scientific community inspiration, knowledge, quality and heritage) • Good content – could be presented better discipline in years to come. Many 6th form and under each of these at least four designs students haven’t even heard of the word relating to the particular title. Two concepts physiology (despite physiology forming a large the logo and strap line, many new materials will were selected for further development and filter through our various channels of these were sent to Council for comment. communication, so all I can really say is ‘watch this space’. Making a final decision on which logo/concept should represent The Society was not easy. Our new look will be launched early in 2007 in Although there was a significant majority of parallel with our new interactive web site. If you votes for the concept we selected, and indeed have any comments either now or when you for the colour palette, trying to get everyone to see everything in place please feel free to agree on miniscule details like the final shape contact me ([email protected]). and position of text on the logo proved challenging, as, at the end of the day, everyone Donna Brown sees things differently and it would appear most people (particularly Exec members) think Copy for the Spring 2007 issue of they have a flare for creativity. With guidance Physiology News should reach the from 35 and our aims in mind we finally Publications Office by 21 December. selected the logo and strap line you see (left). Please send contributions to Obviously this project hasn’t just been about [email protected]

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Physiology News | No. 65 | Winter 2006 | www.physoc.org BIOSCIENCES FEDERATION PN 49

By the time that you read this we will have submitted our views about new RAE metrics to the Department for The BSF has grown in strength during Education and Skills. The BSF the summer months. This is because strongly holds the view that a metrics we have made two important only approach to the RAE after 2008 is appointments to help with the policy wholly undesirable. The Federation work. First, we have recruited Caroline takes the view that metrics should be Wallace. She will have particular there to guide and inform panels but we responsibility for our Animals Science cannot imagine a suitable series of Group and our European Liaison stand alone algorithms for dealing with Group. Caroline has a PhD in all the complexities and different molecular biology and has worked with emphases across the biosciences. We us for the last two years in a also hold the view that metrics should contract/consultancy role. The second not only be about inputs (for example, appointee is Richard Bateman. He has grant income) but also about outputs resigned from a senior position at The (for example, citations). However, the Natural History Museum to become our key element is that metrics are assessed Head of Policy in a part-time capacity. by people and not software. With his background in systematics and plant science he will increase the width Emma Southern, Head of Communications (top) and How do we undertake these policy of our ‘in house’ skill base. These Caroline Wallace, Policy Co-ordinator (above), who reviews? From this summer we have important appointments have become takes on responsibility for the BSF Animal Science developed a closer relationship with possible because of increased and European Liaison Groups. your Society in order that we might membership and, importantly, a in order to achieve this goal. We sat at work together more effectively on key substantial voluntary increase in the tables of eight and took it in turns to policy issues for the biosciences. As an subscription paid by several member give our answers to the questions. issue comes to the fore, we write to all organisations. As a consequence of Interestingly, the answers reflected a Member Organisations and ask them if these appointments we will be even broad swathe of agreement that both they want to nominate someone to be a more effective than hitherto in reacting translation and incentives were not only member of an ad hoc task force to to Government and other enquiries and desirable but essential. However, we work on our response. Therefore if this initiatives. More significantly, we will did not tackle what I believe to be a sort of work interests you at all – and be able to be more proactive. That is, central concern for the BSF. That is, you have something to say (!) – you we can start to identify initiatives as under which modus vivendi will the should let the Society know. they are born and influence their new joint fund operate? Will it be that gestation, and also give birth to some of the MRC or that of the NHS. We are And finally, are you a postdoc or ourselves. In this context, the BSF will absolutely clear about this question: it graduate student looking for a job? If look to you, the member organisation has to be that of the MRC. We should you are, you should find a new page on and the individual, to help with horizon only give grants for potentially our web site helpful. This page scanning and the identification of areas excellent world class research. If areas provides links with very many of the where we should take the lead. need strengthening we should not sites that you might want to look at for pretend that the science is excellent in job advertisements. If you think that Have you seen our response to the order to make an award. If the country there are important links missing please Cooksey enquiry? If you haven’t, it is needs to strengthen an area then inform Emma Southern, Head of on our web site and it gives you some earmarked funds should be used for this Communications idea about what we are doing for you. explicit purpose. The marriage of ([email protected]). I am sure you know that Cooksey is funds for world class research and concerned with putting the funding for capacity building generally reduces the Richard Dyer NHS Research and Development under integrity of awards for both. Chief Executive Officer the same umbrella organisation as MRC grant awards. Following our Biosciences Federation Careers Conferences 2006 submission, the BSF was invited to a University of Reading meeting with the Cooksey team to Saturday 2 December discuss four questions. In summary, For undergraduates, graduates, postgraduates and postdocs from all branches of these can be distilled down to two life science points. They were about translation (the conversion of world class science £12, including refreshments and lunch to medicines and improved clinical For full details of the programme and a booking form, please visit practice) and the incentives to offer http://www.bsf.ac.uk/careers/careersconf.htm scientists, departments and universities

Physiology News | No. 65 | Winter 2006 | www.physoc.org 50 PN UNBELIEVABLE!

cures for human diseases and lives in a university, then any attempt to kill it involves so much time-wasting drivel.

Mice attract two types of unsavoury creatures to your laboratory – animal rights lunatics and the far more Long and tortuous fanatically unbalanced Home Office goodbye to Stud inspector. The former simply has a balanced aura and an infantile ideology Muffin No 1 to maintain. The latter is a humourless bureaucratic equivalent of Hezbollah Cats and Rentokil have it easy. The day I see a mouse pay National with a £60,000 salary and a mortgage Biomedical researchers, however, suffer Insurance contributions I might delude to defend. Whenever a Home Office greatly when they have to kill mice in myself that it is a sentient creature. But inspector turns up, everything in the the lab. it is not. No research has ever workplace grinds to a halt. Experiments demonstrated that rodents think and get cancelled and all sorts of lunacy As a biomedical researcher I am experience suffering in a manner that occurs. afflicted with a serious problem. It's a even closely resembles the human transgenic mouse that has been condition. People who convince In the last place I worked, a researcher engineered with the ability to wreck my themselves otherwise generally have was reprimanded for using secret codes career. I would like to kill it, but the something missing from their lives – on a cage label. Her crime was to write multiple sheets of paper I would have worrying about the non-existent the symbol ®. This Masonic symbol to fill in, the computer records I would consciousness of mice is probably the stood for Rachel. Another was need to input and the bizarre nearest thing to a meaningful admonished for calling a mouse ‘Stud procedures I would be required to relationship that they have. Muffin No 1’ (he produced lots of follow might take me several hours. I kids). The Home Office inspector was just don't have the time. This devious The bottom line is that mice are appalled, though it may have done creature has entered into a conspiracy vermin. Nobody really cares what wonders for the mouse's ego. with the British state, and it has nearly happens to them – even the ‘vegan’ cat as many rights as me. living with the animal rights nutcase Still, there is hope. Someone has down the road eats 200 a year. They devised a way of inserting a In the past 5 years there has been an make small, furry and delicious entrées luminescent gene into mice. They are a explosion of nit-picking legislation for every predator on the planet. genius. If I had my way, the gene intended to provide a paper trail so that would be inserted into every mouse on some faceless bureaucrat can In one study, Britain's domestic cats the planet so that the next time you microscopically follow every stage of were thought to account for more than walked into the kitchen at 2 a.m. you every transgenic mouse's life. 300 million a year. I am surprised the wouldn't need to switch on the light. Home Office hasn't tried to regulate The mice could just be sitting there in No area of transgenic mouse life is that – it's an opportunity to collect an the moonlight innocently fluorescing at sacrosanct. Even if I place a male extra 900 million sheets of paper and each other. And that would make them mouse with a female mouse ‘to do what they could prosecute the odd moggy for far easier to kill. comes naturally’, it is now a procedure filling out the forms wrong. and needs to be recorded first, both on The author would like to remain paper (3 sheets of) and on the If you switched on the kitchen light at anonymous. The last time he killed a computer. Imagine trying that in your 2 a.m. and found one scurrying across mouse in his kitchen he used a £2 trap own bedroom. the floor, you would simply call from B&Q. When he killed a mouse at Rentokil to administer an exotic­ university he filled in three sheets of This bureaucracy wastes time and flavoured warfarin overdose. But if the paper and one Excel file, and needed a taxpayers' money, creates paranoia and mouse is a cute white colour, doesn't training certificate, a named day-to-day can result in a sinister kind of predatory steal your cornflakes, is used to find care officer and a special white suit. wildlife called a Home Office inspector hanging around your workplace. Keith Cormorant Nobody appears to be able to stand up to this regulatory excess. I suspect that This article first appeared in The Times by making animal experimentation as Higher Education Supplement long-winded and tortuous as possible, (http://www.thes.co.uk) on 22 September the hope is to make it difficult to do 2006, and is reprinted here with any real work. permission.

Physiology News | No. 65 | Winter 2006 | www.physoc.org OBITUARIES PN 51

Douglas L Ingram Physiology, University of California at first at the ARC Hannah Research Davis, USA (1968–1969), Lecturer in Institute (1958–1961) and then at The 1929–2006 Climatic Physiology, Department of Babraham Institute, Cambridge. He Biological Anthropology, University of carried out many detailed Cambridge (1974–1997), Special investigations, often with co-workers, Professor of Environmental Physiology, into the role of thermosensitive regions University of Nottingham (1981–1989), deep in the body and on the skin and Supervisor in Physiology surface. His research was concerned (1969–1999) for undergraduates in with both behavioural and autonomic Medical, Veterinary and Natural systems, such as blood flow and Sciences at Gonville and Caius College, metabolism, and it was concluded that Newnham College, Churchill College the control systems depend on the and Christ’s College, University of integration of signals from all over the Cambridge. He was a Member of the body. At this stage he also initiated High Table at Christ’s until March studies into interactions between 2006. nutrition and the thermal environment. The effects of changes in environmental Doug was always passionate about conditions were investigated in relation biology. He was born and brought up in to a range of variables such as sensible Birmingham where the parks and small and evaporative heat loss and the rivers provided early opportunities for quantity of food eaten, and discoveries about the natural demonstrated a striking interaction environment. He shared with his father, between behavioural thermoregulation The death of Doug Ingram from cancer a shoe-repairer, the intellectual and food intake. at his home in Whittlesford, Cambridge curiosity and practical ability that are on 21 March marks the end of a unique essential for success as a first-rate These studies resulted in numerous scientist and human being. He had a research scientist. Enlightened post-war publications in learned journals, tremendous love of science and lived policies on further education for those including The Journal of Physiology, life to the full. His research career with academic ability but limited and joint publication of the definitive spanned four decades during which financial resources, together with a City book on Man and animals in hot time he made major contributions to the of Birmingham Exhibition, enabled him environments. This work continues to fields of reproductive physiology, to join his local university during a provide insight into fundamental thermal and environmental physiology, period of immense research mechanisms of thermoregulation. In the and nutritional and developmental achievement and leadership. He was an current environment of global climate physiology. Not only did his findings undergraduate in the Department of change his contribution is particularly result in significant insight into Zoology under Peter Medawar and then relevant to an understanding of the fundamental biology but they also a PhD student in the Department of potential for homeostatic adaptation in paved the way for current applications Anatomy under Solly Zuckerman. His all mammalian species. In recognition in biomedicine. He was extremely studies on reproductive physiology of his achievements, he was awarded modest about his ability and were interrupted by a 2 year period of Individual Merit Promotion to Senior achievements, frequently attributing National Service as a Royal Air Force Principal Scientific Officer in 1975, and major advances to his colleagues. Education Officer, after which time he a DSc by the University of Birmingham Despite being a full-time research was awarded a 3 year MRC Research and elected a Fellow of the Institute of scientist, Doug also made time to help Fellowship to return to the University Biology in 1976. others – his knowledge and enthusiasm of Birmingham under Zuckerman. His for science have been passed on to work was concerned with the problem The last 15 years of Doug's research numerous students and colleagues of atresia which causes a decline in the career saw a remarkable period of world-wide. number of oocytes in the mammalian scientific achievement. He did not seek ovary. It provides the impetus for many high office but chose to continue in The major part of Doug’s research current studies on germline stem cells practical research, during which time he achievement occurred between 1961 and follicular renewal in the postnatal demonstrated unique ability as an and 1989 at the ARC Institute of mammalian ovary; studies that have integrative scientist. This research, with Animal Physiology, Cambridge/BBSRC significant clinical implications with many colleagues, visiting scientists and Babraham Institute, in the Departments respect to ovarian failure. students, highlighted his continuing of Applied Biology, Cell Biology, and flair for asking the right questions and Molecular and Cellular Physiology. He Key aspects of environmental the ability to answer them by was a Member of The Physiological physiology, especially thermoregulation developing new techniques. Of Society from 1963 to 2001. Doug was and adaptation to the thermal particular importance are studies on the Visiting Lecturer and Research Fellow, environment, formed the basis for the influence of environmental temperature Departments of Veterinary and Medical major phase of Doug's research career, and energy intake on postnatal

Physiology News | No. 65 | Winter 2006 | www.physoc.org 52 PN OBITUARIES development. Some environmentally exciting and comprehensible. advice and help on numerous and nutritionally induced changes were Especially revealing to my mind is the occasions’, ‘we all liked Doug so found to be at least as great as those extraordinary phenotypic change much, and his quiet modesty belied his determined by genetics and the brought about by ambient temperature considerable achievements and underlying mechanisms involved differentials, surely a timely leveller for international reputation in the scientific physiological, biochemical, cellular and the almighty gene.’ field.’ Perhaps most of all Doug was a molecular modifications. His studies person of wonderful humanity and highlighted the role of the endocrine Following formal retirement, Doug sense of fun. He is greatly missed and system, especially thyroid and growth continued to demonstrate his love of will not be forgotten; his legacy lives hormones, in these responses. For science and desire to seek answers to on through the wisdom and kindness he example, it was found that temperature the deeper questions about life. In gave to his family, colleagues and and energy intake influence thyroid addition to teaching undergraduates, he friends. hormone metabolism through separate read widely and studied astronomy and mechanisms, and that the abundance of cosmology at the Institute of Joy Dauncey nuclear thyroid hormone receptors is Continuing Education, University of Fellow of Wolfson College, University of Cambridge determined by an interaction between Cambridge. In his final year he showed thermal and nutritional factors. This tremendous courage in the face of MOLECULAR TECHNIQUES deteriorating health. He continued to research provided new insight into FOR LIFE SCIENCES fundamental problems of growth and pursue his interests in philosophy, development and has led to increased baroque music, sailing and walking, PCR Theory and Practice understanding of the mechanisms by and enjoyed intellectual discussions 22–26 January 2007 which early environment has both with former colleagues and friends. A A five day course to introduce immediate and long-term effects on testimonial in 1955 stated ‘He is a man participants to this core technique health and disease. When Doug retired of high intelligence, bright disposition covering the basics to quantitative from The Babraham Institute, the of ideas and practical ability. His moral Real-time PCR. Cost: £870 Director wrote about his impressive character is of the highest standard’. (Standard), £740 (CPD Accredited). productivity and pioneering research Numerous letters of condolence to his For further information and since Individual Merit Promotion, and family provide continuing testimony: ‘a application form visit said ‘As the march of science has been truly good and kind man’, ‘I remember www.caledonian.ac.uk/mtls taking an ever more molecular path, it him with great affection as a boss who has been reassuring for me to know that actually cared about those under his IUPS some of our staff have continued a charge – he was tolerant, understanding Kyoto, Japan (27 July–1 August whole animal approach. In this regard I and inspirational’, ‘a wonderful 2009) http://www.iups.org have always found your work both colleague and I always appreciated his

Physiology News | No. 65 | Winter 2006 | www.physoc.org IMAGES OF BRAZIL Photos from the sessions at the Joint International Meeting with the Brazilian Physiological Society will be published in PN 66

(photos by Prem Kumar and Nana Voitenko)