1329

W-

i

Fig. 2-Case 5: S.E.M. Presumably neoplastic large cell of H.D.with attached small lymphocytes.The large cell displays many surface projections of variable

lengths and complexity. Some projections show terminaldilatation. x 5200.

cessed non-neoplastic conditions of the lymphoid tissue orin non-Hodgkin lymphomas. The architectural featuresof the large cells, although unique in our experience, donot shed light on the origin of the neoplastic cells in H.D.

Hematopathology Section,Laboratory of Pathology,National Cancer Institute,

National Institutes of Health,Bethesda, Maryland 20014, U.S.A.

R. C. BRAYLANE. S. JAFFEC. W. BERARD.

ENHANCEMENT OF LEVODOPA-INDUCED

GROWTH-HORMONE STIMULATION BY

PRACTOLOL

SIR,-Massara and Camanni reported that propranololstrikingly enhanced the growth-hormone response to

levodopa 1 ; others have reported similar findings with othergrowth-hormone-secretion-evoking stimuli. 2,3 Moreover,the therapeutic effect of levodopa on tremor, in Parkin-sonism, can be ameliorated in many cases by propranolol

PLASMA-GROWTH-HORMONE LEVELS IN NORMAL AND OBESE

SUBJECTS AFTER LEVODOPA AND BETA-BLOCKERS

* Irrespective of time of occurrence.t p < 005.t < 001.

and other beta-blockers, provided that they can cross theblood/brain barrier; practolol does not penetrate thisbarrier at all in man, and only slightly in animals, and istherapeutically ineffective. 4,5

Besides confirming the potentiating effect of propranololon growth-hormone response to levodopa in normal people, 6

1. Camanni, F., Massara, F. Lancet, 1974, i, 942.2. Blackard, W. G., Heidingsfelder, S. A. J. clin. Invest. 1968, 47, 1047.3. Parra, A., Schultz, R. B., Foley, T. P., Jr., Blizzard, R. M. J. clin.

Endocr. Metab. 1970, 30, 134.4. Kissel, P., Tridon, P., Andr&eacute;, J. M. Lancet, 1974, i, 403.5. Scales, B., Cosgrove, M. B. J. Pharmac. exp. Ther. 1970, 175, 338.6. Delitala, G., Masala, A., Alagna, S., Devilla, L. Studi Sassar. 1974,

52, 1.

we have demonstrated the same effect in obese subjectsand have shown that the increase in plasma-growth-hormone after levodopa and practolol is equal to or evenhigher than that after levodopa and propranolol, both innormal and obese subjects (see accompanying table).Since practolol cannot penetrate the blood/brain barrier inman, the mechanism responsible for potentiating the

growth-hormone response to levodopa may be peripheral.Cattedra di Medicina Costituzionale

ed Endocrinologia,Universit&agrave; di Sassari,

Viale S. Pietro 12, 07100 Sassari,Sardinia, Italy.

G. LOTTIG. DELITALAA. MASALA.

ANTI-INFLAMMATORY DRUGS ANDTISSUE COPPER

SiR,&mdash;Miss Forder (Oct. 26, p. 1009) points out thatcopper may have an important role in the biosynthesis ofprostaglandins in the endometrium. I should like to takethis suggestion further by indicating the possible importanceof copper in the action of anti-inflammatory drugs and alsoin the action of penicillamine in rheumatoid arthritis andscleroderma.The inflammatory process in several tissues is associated

with an increase in the extractable prostaglandin content ofthe tissue involved,! and Greaves et awl. found a mixtureof prostaglandins E and F in the perfusate obtained frominflamed skin. The E series of prostaglandins have moreinflammatory activity as judged by the reaction of skin toan intradermal injection than the F series, and pre-treat-ment with low doses of p.G.F2 suppresses the inflammatoryresponse to injury in rats,3 thus suggesting that a roughdivision into active inflammatory E series and anti-inflam-matory F series can be postulated. The shift from p.G.E2synthesis to P.G.F2 due to copper inhibition of P.G.E2synthetase described by Maddox 4 could be expected toreduce inflammation in a tissue. Maddox also reportedthat penicillamine, indomethacin, fenclozic acid, phenyl-butazone, and salicylic acid all accelerated the inactivationof P.G.E2 synthetase in vitro. In addition to having anti-arthritic actions all these drugs will chelate copper.Work linking copper, inflammation, and analgesics has

been done by Sorenson,5 who demonstrated that copperacetate alone has an anti-inflammatory action in an experi-mental model of inflammation and suggested that the activemetabolite in anti-inflammatory activity is a copperchelate. He tested this hypothesis firstly by preparingcopper chelates of compounds not possessing anti-inflam-matory properties and finding that copper chelates ofanthranilic acid and 3,5,di-isopropylsalicylic acid wereactive in reducing inflammation, and secondly by findingthat copper chelates of aspirin, UP 83 (an analogue offlufenamic acid), penicillamine, and P.A.T. (1-phenyl-5-aminotetrazol) had up to 20 times the anti-inflammatoryactivity of the basic compound. These chelates also showedanti-ulcer activity in animal experiments and inhibited thesecretion of acid and pepsin in the stomach.He postulated that the anti-inflammatory activity of a

drug is mediated through the copper chelate of that drug,and that the chelate is formed by the drug combining withtissue copper. This combination with tissue copper couldlead to a relative lack of free copper ions in the tissues, andin susceptible tissues like the stomach this could resultin more synthesis of the P.G.E series and less synthesis ofthe P.G.F series, leading to ulceration. This undesirable

1. Vane, J. R. Nature New Biol. 1971, 231, 232.2. Greaves, M. W., S&oslash;ndergaard, J., McDonald-Gibson, W. Br.

med. J. 1971, ii, 258.3. Cuthbert, M. F. in The Prostaglandins; p. 97. London, 1973.4. Maddox, I. S. Biochim. biophys. Acta, 1973, 306, 74.5. Sorenson, J. R. J. Proceedings of Medicinal Chemistry Section,

American Chemical Society (in the press).

1330

side-effect could be mitigated by administration of theanti-inflammatory drug as a copper chelate, and, if anti-inflammatory activity is in fact dependent on the presenceof a copper chelate acting on prostaglandin synthesis, thiswould considerably enhance the efficacy of the drug.

In addition to being of therapeutic value in the treatmentof rheumatoid arthritis, the copper-chelating drug D-

penicillamine has also been used in the treatment of

scleroderma, a condition in which there is a higher propor-tion than normal of reducible aldimine cross-linkages incollagen. Penicillamine therapy results in a decrease in theproportion of reducible cross-linkages in skin from activelesions 6 and part of this action may be due to inhibitionof the copper-containing enzyme lysine oxidase by chela-tion of copper in the tissues although this probably onlyoccurs at very high doses of penicillamine.

In hepatic cirrhosis where there is an increased collagencontent of the liver due- to the associated fibrosis, there areabnormally high copper concentrations. 7 Penicillamine

therapy in chronic hepatitis and cirrhosis has been usedwith benefit after the report by Ruiz-Torres 11 that penicil-lamine prevented experimentally induced collagenisationof the liver and also increased the solubility of the collagen.The question is how much of this anti-collagen activity isdue to the chelating action of the penicillamine, resulting inremoval or binding of the excess copper ? If that is the ex-

planation, then further devices to remove excess coppercould be adopted, such as the administration of zinc com-pounds which compete with copper for absorption fromthe small intestine.9

It seems possible that the anti-inflammatory action ofanalgesics, such as aspirin, and the anti-inflammatoryaction and also the anti-collagen effect of penicillamine,may all be linked by their effect on tissue copper, and thatcollaboration between workers in trace-element meta-

bolism and those in experimental therapeutics would provefruitful.

Institute of Clinical Science,Grosvenor Road,Belfast BT12 6BJ. MARGARET E. ELMES.

CYCLIC A.M.P., SUBSTANCE P, ANDMORPHINE ANALGESIA

SIR Dr Brammer and Dr Paul (Nov. 2, p. 1084)describe stimulation of cycliC-A.M.P. formation by morphinein midbrain thalamus. This raises the possibility that cyclicA.M.P. mediates the action not only of physiologically activemolecules like hormones and putative neurotransmittersbut also of pharmacologically active substances.We have also shown that substance P, a brain peptide

and an antagonist of morphine analgesia," also stimulatescyclic-A.M.P. formation (by 52%, P<0.05) in the humanthalamus in vitro. Thus it seems that both morphine andsubstance P may act via cyclic A.M.P. Therefore the actionof substance P in antagonising morphine analgesia may be toinhibit the binding of morphine to its specific receptor.

Finally, one must be very cautious about interpreting thephysiological importance of stimulation of cyclic-A.M.P.formation in brain by any compound, since molecules of asdifferent structures as Na F, adenosine, substance P,prostaglandins, and catecholamines all stimulate cyclic-A.M.P. formation in brain.

Radioisotope Department,St. Vincent’s Hospital, Dublin 4,and Biochemistry Department,

Trinity College, Dublin 2.M. J. DUFFYD. POWELL.

6. Herbert, C. M., Lindberg, K. A., Jayson, M. I. V., Bailey, A. J.Lancet, 1974, i, 187.

7. Underwood, E. J. in Trace Elements in Human and Animal Nutri-tion; p. 67. New York, 1971.

8. Ruiz-Torres, A. Arzneimittel-Forsch. 1968, 18, 594.9. Underwood, E. J. in Trace Elements in Human and Animal Nutri-

tion; p. 218. New York, 1971.10. Stern, P., Hadzovic, J. Archs int. Pharmacodyn. 1973, 202, 259.

VIRAL-CODED INFORMATION IN HUMANRHABDOMYOSARCOMA CELLS

SIR,&mdash;We have lately reported the transformation ofhuman skin fibroblasts by D.N.A. extracted from a humanrhabdomyosarcoma cell-line. 1 own the basis of these findingswe speculated that the D.N.A. which transformed the fibro-blasts contained oncorna viral-coded information. Weshould now like to report the results of studies to investi-

gate further the possibility that these human rhabdomyo-sarcoma cells contain viral-coded information responsiblefor their malignant state.The cultured rhabdomyosarcoma cells were very pleo-

morphic and had the growth characteristics of cancer cells(they grew in multilayers, and on top of normal cells).It is therefore reasonable to assume that these cells re-presented malignant tissue rather than an outgrowth offibroblasts. Numerous giant cells were present in the

Part of the cytoplasm of cultured rhabdomyosarcoma cells

( x 78,000).Several virus-like particles are shown within the cytoplasmic

matrix.

culture; some of these contained several nuclei of differentsize and shape, and it is possible that they may reflect thepresence of unstable integrated viral genome. Furtherevidence for the presence of incomplete viral informationis provided by the finding of abnormal mitotic figurescontaining damaged chromosomes. We have found similarchromosomal abnormalities in a human cell-line trans-

formed by Rous sarcoma virus (unpublished), as well as ina variety of mammalian cells known to contain incompletesarcoma viral genomes .2,3 In addition, electron-microscopicexamination of the cultured rhabdomyosarcoma cellsrevealed the presence of electron-dense structures whichmight represent the morphological expression of incompleteC-type virus (see figure). In addition, we found that acytoplasmic fraction of the rhabdomyosarcoma contained a

- high level of activity of the viral-coded enzyme, R.N.A.-directed D.N.A. polymerase. In general, this viral-codedenzyme is only associated with oncorna viruses and initiatesthe synthesis of a D.N.A. copy of the viral R.N.A. which thenintegrates into, and replicates within, the affected cells,thereby maintaining their malignant stated 4 Therefore,the demonstration of this enzyme provides importantadditional evidence in support of the possible role of onco-

1. Karpas, A., Tuckerman, E. Lancet, 1974, i, 1138.2. Karpas, A., Cawley, J., Tuckerman, E., Flemans, R., Hayhoe,

F. G. J. Br. J. Cancer, 1971, 25, 779.3. Karpas, A., Cawley, J., Tuckerman, E. Z. Krebsforsch. 1972, 78, 51.4. Temin, H. M. Ann. Rev. Microbiol. 1971, 25, 609.