- Email: [email protected]
AGGRESSION AND THE YY SYNDROME
355 in vitro are apparently a prerequisite for cellular maturation both functionally and morphogenetically. The fact that also certain cellular fractions (fig. 2) induce in-vitro hybridisation between lymphocytes, might implicate cell fractions of lymphocytes of individuals treated with certain drugs," or subjected to certain infections or irradiations, in the genesis of some malignancies or autoimmune diseases-presumably as a consequence of lethal hybridisation of lymphocytes by other lymphocytes, or of lymphocytes and other target tissues. unit wishes to acknowledge the help of grant no. 4X-2507 from the National Institutes of Health, Bethesda, U.S.A., and that of the Association of Aid For Crippled Children, New York, U.S.A. Medical Genetics Unit, Ain-Shams University, NEMAT HASHEM. Cairo, Egypt, U.A.R.
suspected case. Most of the investigated sera were taken from blood-donors who have given blood for many years, and there have never been any symptoms or signs of transfusion hepatitis among the recipients. The high percentage of H.l.M.-positive blood-donors in our investigation makes it practically impossible to use only H.I.M.-negative individuals as blood-donors. J. HOLMGREN O. MÖLLER and Bacteriological Laboratory the Central Blood Bank, Borås. J. STANEK. a
I This
AGGRESSION AND THE YY SYNDROME
SiR,łThere is a growing number of reports about men with an XYY sex-chromosome complement, and the following case presenting as an acute medical emergency seems worthy of record.
A PREREGISTRATION PROBLEM SiR,łThere must be many women who face the prospect of registering while they have a young family to care for, and who find it harder to solve than Dr. Forrester (July 6, p. 51) implies in his reply to Professor Brooke (June 29, p. 1425). Together with several of my colleagues-recently qualified, unregistered mothers-I believe that the problem is not so much the place of residence as the hours of work. It is simply impossible to do a full-time house job and to spend a One or reasonable amount of time with one’s family. the other must suffer. It seems to us that some form of part-time house job is the only solution, and that, with good will and good planning, it can be organised without at all lowering the standard of care. At present this could be done only by ad-hoc arrangement with particularly cooperative hospitals, but we hope one day to see it become an acceptable and regular way of fulfilling in these circumstances the requirements for registration. CATHERINE OPPENHEIMER. Oxford.
HEPATITIS/INFECTIOUS-MONONUCLEOSIS ANTIBODY TEST
SiR,łThe hepatitis/infectious-mononucleosis (H.I.M.) antibody test has been recommended 11 as a useful serological test for the confirmation of a tentative clinical diagnosis of viral hepatitis or infectious mononucleosis. Its use has also been proposed for studies of H.l.M. antibodies in blood-donors and the incidence of viral hepatitis in their recipients. In our laboratory we have studied the prevalence of a positive H.I.M. test in healthy blood-donors in the region of Boras, a Swedish inland town with about 75,000 inhabitants. Blood-sera from 296 healthy blood-donors were tested for H.l.M. antibodies. These donors had never had any clinical signs of hepatitis or infectious mononucleosis, had not been abroad during the past eight months, and had a serum-glutamicpyruvic-transaminase value less than 30 units. Blood-sera from 11 patients with high Paul-Bunnell titres (more than 256) were also investigated for H.l.M. antibodies. The H.I.M. test was performed as described by the manufacturer (Bolin Laboratories, Inc., Glendale, Arizona) and positive and negative control sera were always included. Of the 296 blood-donors, 189 (64-0%) had demonstrable H.l.M. antibodies in their blood-sera. When recording the blood-donors according to their ABO and rhesus blood-groups, the percentage of H.I.M.-positive individuals was found to be about the same (64+-2%) in the groups. Of the 11 blood-sera which gave Paul-Bunnell titres greater than 256, 7 (64%) gave a positive H.l.M. test, and 4 were negative. A test which is positive in 64% of healthy blood-donors is obviously of little value to confirm the diagnosis of hepatitis in 10.
G. J., Kadaghidze, Z. G., Sura, S. N. Lancet, 1968, i. 148. 11. Bolin, V. S., Chase, B. S., Alsever, J. B., Mann, F. D. Fedn Proc. 1965,
Svet-Moldavsky, 24, 159.
A 23-year-old private in a Guards regiment was drinking with friends and after a moderate quantity of alcohol complained of a left-sided chest pain which he described as sharp in quality, lasting 2-3 minutes at a time, and recurring at intervals of 10 minutes or so. He had experienced this pain intermittently for 4-5 years and twice during the previous 4 months. Following the onset of the pain on this occasion he became very violent, striking anybody in sight, particularly when they tried to restrain him. He was admitted to the casualty department where four porters had difficulty in holding him down. When liberated he was docile and
cooperative. As might be expected of a Guardsman the patient was above average in height (6 ft. 2 in., 188 cm.), and was well built. On clinical examination, X-ray of the chest, and electrocardiographic examination no cause could be found for the pains, which did not recur in hospital. His violent behaviour was not without precedent, and he described an unfortunate liability to get into fights. He said that he easily took offence and that his normal reaction was to hit first and ask questions afterwards. The patient was the second in a sibship of two. He had passed the developmental milestones at the expected ages. According to the parents he had been a quiet child, and not aggressive; he had never been involved in fights at school, which he had attended regularly, but he had been inclined to be destructive. At the age of 12 years he had had a minor road accident causing lacerations to the face and after this his parents noticed a striking change in his behaviour. He became resentful and would not tolerate anyone trying to push him around, and would not stand any criticisms of himself or his friends. He spent a lot of time on his own and would at times work out his feelings by throwing a large knife into a block of wood. After leaving school he had many changes of job, sometimes staying in one place of work for only 2-3 days. He had made two attempts to join the army, being turned down on the first occasion because of bed-wetting. During the 8 months that he had been in the army he had on several occasions " gone berserk " in the barrack-hut, and as a result of his violence several of his fellow soldiers had had to go on sick-
parade. When the patient was interviewed in hospital there was no evidence of any serious psychiatric illness. He was pleasant and cooperative, did not appear depressed or psychotic, and On the basis of the was not at all aggressive or paranoid. hospital interviews and his behavioural history a personality disorder of a somewhat immature, aggressive, and rather dependent type was diagnosed. It was felt, however, that the possibility of temporal-lobe epilepsy could not be excluded. Unfortunately the patient did not keep an appointment for an electroencephalogram. Because of the combination of a personality disorder and his above-average height 12 the peripheral-blood leucocytes were karyotyped. Fifty cells were counted and analysed and twenty-six had 46 chromosomes with a normal sex-chromosome constitution, while nineteen 1. 2.
Price, W. H., Whatmore, P. B. Br. med. J. i, 533. Jacobs, P. A., Jacobs, A., Price, W. H., Court-Brown, W. M., Brittain, R. P., Whatmore, P. B. Ann. hum. Genet. 1968, 32, 339.
356 had 47 chromosomes and an XYY sex-chromosome constitution. The remaining five cells did not have a consistent analysis. The patient was reported as a 46, XY/47, XYY mosaic.
grateful to Dr. Patricia Jacobs for the report on the chromoconstitution, to Dr. W. H. Price for his advice, and to Mr. lain Paterson, psychiatric social worker, for information derived from the patient’s parents. M. B. MATTHEWS Western General Hospital, P. W. BROOKS. Edinburgh 4. We are
the variants of glucose-6-phosphate dehydrogenase), so that temporary nomenclature could be suggested. Laboratoire d’Enzymologie, Unité de Recherches Isotopiques de l’I.N.S.E.R.M., P. BOIVIN C. GALAND. Hôpital Beaujon, 92 Clichy, France.
a
some
ANTILYMPHOCYTE SERUM SIR,-Your annotation (July 27, p. 204) does less than justice to this important subject. The discovery that " antilymphocyteserum products may abolish the immunological opposition to grafts " is at least five years old, and even if you mean heterografts the discovery goes back to 19661 or even earlier. The effectiveness and advantages of IgG prepared from antilymphocyte serum (A.L.S.), instead of crude serum, have been stressed in many papers over the last few years from our own laboratory and elsewhere, including one in The Lancet,2 and a great deal of work has been done which throws light on the mode of action of A.L.S. in vivo and in vitro. We realise that space is limited but feel that you might have helped those of your readers who are unfamiliar with this work to gain a fuller and more balanced picture by citing one of two recent review articles. One such appeared last year3 and a fuller and more up-to-date review from the same author is to appear shortly.4 University Department Medical
of
Surgical Science,
School,
Edinburgh,
8.
7,X DA M. F. A WOODRUFF KEITH JAMES.
*** Not many months ago we had a leader5 on antilymphocyte serum, so our annotation simply aimed to pick up a few of the latest findings, of which there are a vast number.-ED. L.
VARIANTS OF ERYTHROCYTIC PYRUVATE KINASE SIR,-Professor Lohr and his colleagues 6have reported electrophoretic abnormalities of erythrocytic pyruvate kinase (P.K.) in hxmolysates of patients with congenital hxmolytic anxmia. In one of the cases described the Michaelis constant of the enzyme for phosphoenolpyruvate (P.E.P. Km) was high.’ We have studiedcertain characteristics of concentrated P.K., extracted from hxmolysates of ten normal subjects, seven patients with congenital hxmolytic non-spherocytic anxmia with homozygous deficiency in P.K., and six collateral relatives in whom the deficiency was present in a " heterozygous " state. Of the seven homozygous subjects, in three the P.E.P. Km was equal to that of normal subjects; in two, it was about a quarter of normal; and in two it was considerably increased, as in the case described by Lohr.’ In five of the six heterozygous subjects it was normal; in one it was moderately increased. The Michaelis constant for adenosine phosphate in three patients with congenital hxmolytic anaemia with P.K. deficiency was found to be equal to that of control subjects. Such findings confirm the existence of various types of molecular abnormalities in connection with P.K. deficiency. It therefore seems desirable that criteria should now be defined for characterisation of these various types (as has been done for 1. Monaco, A. P., Wood, M. L., Gray, J. G., Russell, P. S. J. Immun. 1966, 96, 229. 2. Anderson, N. F., James, K., Woodruff, M. F. A. Lancet, 1967, i, 1126. 3. James, K. Clin exp. Immun. 1967, 2, 615. 4. James, K. Progr. Surg. (in the press). 5. Lancet, 1967, ii, 661. 6. Blume, K. G., Löhr, G. W., Rüdiger, H. W., Schalhorn, A. ibid. 1968,
i, 529. 7. Löhr, G. W., Blume, K. G., Rüdiger, H. W., Sokal, G., Gulbis, E. ibid. p. 753. 8. Boivin, P., Galand, C. Nouv. Revue fr. Hémat. 1968, 8, 201.
ALKALINE-PHOSPHATASE-SECRETING TUMOUR OF LUNG
SIR,- There have been several reports of tumours not involving bone, with a raised serum-alkaline-phosphatase. 12 The importance of these observations lies in the fact that the neoitself may be the source of the alkaline phosphatase. It is therefore important to characterise the different types of tumour alkaline phosphatase in order to determine whether there is any characteristic which is typical of any particular type of tumour. I have lately had a patient whose carcinoma of the lung contained a high concentration of alkaline phosphatase and who also had a raised level of enzyme in the serum. The patient was a man, aged 73 years, who presented at the
plasm
Manchester Royal Infirmary with a 6-month history of weight loss, hxmoptysis, and dyspnoea. Chest X-ray suggested a neoplasm in the left upper-lobe bronchus. Investigations were limited in view of the patient’s age and physical state, but he was found to have a serumalkaline-phosphatase of 20 King-Armstong units on three occasions. He died a month after admission. Necropsy showed an infiltrating carcinoma approximately 4-0 cm. in diameter in the left upper-lobe bronchus. Secondary deposits were present in the left kidney, both adrenal glands, and the para-aortic, cervical, and right axillary lymphnodes, and a small solitary deposit was present in the left lobe of the liver. There were no detectable bone secondaries, and the bileduct was patent. Histologically the tumour was a moderately welldifferentiated squamous-cell carcinoma. Cryostat sections were cut from both theprimary tumour and the secondary deposits, and the sections were stained for alkaline phosphatase by the method of Burstone.3 Both stained strongly. Alkaline phosphatase was extracted from the tumour with butanol,4 and disc electrophoresis was carried out using 5-0% acrylamide gel in a Shandon electrophoresis tank. The enzyme extracted from the tumour was run in parallel with a sample of serum from a case of bone disease (Paget’s disease), and the gels were stained in the same "
"
medium as was used for the cryostat sections. The tumour extract showed two bands: a strong band moving slightly more slowly than the fastest band in the serum, and a weaker band nearer the origin. The effect of heat on the enzyme was determined by removing samples at intervals from tumour extract incubated at 56°C. The amount of enzyme left in each sample was determined by the department of biochemistry at Manchester Royal Infirmary. The results were as follows:
There was 66% inhibition at 10 minutes and 83% at 30 minutes, after which there was no further inhibition. Electrophoresis of a sample after heat inactivation showed that all the remaining activity was in the broad fast band. No inactivation of enzyme activity was seen in the presence of 0-005 M L-phenylalanine. The idea that a tumour may be the source of a raised serum-
alkaline-phosphatase has been put forward before, but so far there are few studies of the biochemical properties of the types of alkaline phosphatase found in tumours. Fishman et a1.2 reported a case of a patient with carcinoma of the lung with metastases in lymph-nodes and other organs in which both the tumour and serum showed an atypical alkaline phosphatase in that it contained a very heat-stable L-phenylalanine-sensitive enzyme which accounted for 50% of the total serum activity. The enzyme from the present tumour differs from this in that it was largely heat-labile and insensitive to L-phenylalanine. Considerably more work will need to be done before any pattern can be associated with any particular tumour type. Department of Pathology, University, Manchester 13.
The
W. R. TIMPERLEY.
1. Schlang, H. A., McHenry, L. E., Jr. N.Y. State J. Med. 1962, 62, 3811. 2. Fishman, W. H., Inglis, Norma I., Stolbach, L. L., Krant, M. J. Cancer Res. 1968, 28, 150. 3. Burstone, M. S. J. natn. Cancer Inst. 1958, 20, 601; J. Histochem. Cytochem. 1958, 6, 322. 4. Morton, R. K. Biochem. J. 1954, 57, 595.
suspected case. Most of the investigated sera were taken from blood-donors who have given blood for many years, and there have never been any symptoms or signs of transfusion hepatitis among the recipients. The high percentage of H.l.M.-positive blood-donors in our investigation makes it practically impossible to use only H.I.M.-negative individuals as blood-donors. J. HOLMGREN O. MÖLLER and Bacteriological Laboratory the Central Blood Bank, Borås. J. STANEK. a
I This
AGGRESSION AND THE YY SYNDROME
SiR,łThere is a growing number of reports about men with an XYY sex-chromosome complement, and the following case presenting as an acute medical emergency seems worthy of record.
A PREREGISTRATION PROBLEM SiR,łThere must be many women who face the prospect of registering while they have a young family to care for, and who find it harder to solve than Dr. Forrester (July 6, p. 51) implies in his reply to Professor Brooke (June 29, p. 1425). Together with several of my colleagues-recently qualified, unregistered mothers-I believe that the problem is not so much the place of residence as the hours of work. It is simply impossible to do a full-time house job and to spend a One or reasonable amount of time with one’s family. the other must suffer. It seems to us that some form of part-time house job is the only solution, and that, with good will and good planning, it can be organised without at all lowering the standard of care. At present this could be done only by ad-hoc arrangement with particularly cooperative hospitals, but we hope one day to see it become an acceptable and regular way of fulfilling in these circumstances the requirements for registration. CATHERINE OPPENHEIMER. Oxford.
HEPATITIS/INFECTIOUS-MONONUCLEOSIS ANTIBODY TEST
SiR,łThe hepatitis/infectious-mononucleosis (H.I.M.) antibody test has been recommended 11 as a useful serological test for the confirmation of a tentative clinical diagnosis of viral hepatitis or infectious mononucleosis. Its use has also been proposed for studies of H.l.M. antibodies in blood-donors and the incidence of viral hepatitis in their recipients. In our laboratory we have studied the prevalence of a positive H.I.M. test in healthy blood-donors in the region of Boras, a Swedish inland town with about 75,000 inhabitants. Blood-sera from 296 healthy blood-donors were tested for H.l.M. antibodies. These donors had never had any clinical signs of hepatitis or infectious mononucleosis, had not been abroad during the past eight months, and had a serum-glutamicpyruvic-transaminase value less than 30 units. Blood-sera from 11 patients with high Paul-Bunnell titres (more than 256) were also investigated for H.l.M. antibodies. The H.I.M. test was performed as described by the manufacturer (Bolin Laboratories, Inc., Glendale, Arizona) and positive and negative control sera were always included. Of the 296 blood-donors, 189 (64-0%) had demonstrable H.l.M. antibodies in their blood-sera. When recording the blood-donors according to their ABO and rhesus blood-groups, the percentage of H.I.M.-positive individuals was found to be about the same (64+-2%) in the groups. Of the 11 blood-sera which gave Paul-Bunnell titres greater than 256, 7 (64%) gave a positive H.l.M. test, and 4 were negative. A test which is positive in 64% of healthy blood-donors is obviously of little value to confirm the diagnosis of hepatitis in 10.
G. J., Kadaghidze, Z. G., Sura, S. N. Lancet, 1968, i. 148. 11. Bolin, V. S., Chase, B. S., Alsever, J. B., Mann, F. D. Fedn Proc. 1965,
Svet-Moldavsky, 24, 159.
A 23-year-old private in a Guards regiment was drinking with friends and after a moderate quantity of alcohol complained of a left-sided chest pain which he described as sharp in quality, lasting 2-3 minutes at a time, and recurring at intervals of 10 minutes or so. He had experienced this pain intermittently for 4-5 years and twice during the previous 4 months. Following the onset of the pain on this occasion he became very violent, striking anybody in sight, particularly when they tried to restrain him. He was admitted to the casualty department where four porters had difficulty in holding him down. When liberated he was docile and
cooperative. As might be expected of a Guardsman the patient was above average in height (6 ft. 2 in., 188 cm.), and was well built. On clinical examination, X-ray of the chest, and electrocardiographic examination no cause could be found for the pains, which did not recur in hospital. His violent behaviour was not without precedent, and he described an unfortunate liability to get into fights. He said that he easily took offence and that his normal reaction was to hit first and ask questions afterwards. The patient was the second in a sibship of two. He had passed the developmental milestones at the expected ages. According to the parents he had been a quiet child, and not aggressive; he had never been involved in fights at school, which he had attended regularly, but he had been inclined to be destructive. At the age of 12 years he had had a minor road accident causing lacerations to the face and after this his parents noticed a striking change in his behaviour. He became resentful and would not tolerate anyone trying to push him around, and would not stand any criticisms of himself or his friends. He spent a lot of time on his own and would at times work out his feelings by throwing a large knife into a block of wood. After leaving school he had many changes of job, sometimes staying in one place of work for only 2-3 days. He had made two attempts to join the army, being turned down on the first occasion because of bed-wetting. During the 8 months that he had been in the army he had on several occasions " gone berserk " in the barrack-hut, and as a result of his violence several of his fellow soldiers had had to go on sick-
parade. When the patient was interviewed in hospital there was no evidence of any serious psychiatric illness. He was pleasant and cooperative, did not appear depressed or psychotic, and On the basis of the was not at all aggressive or paranoid. hospital interviews and his behavioural history a personality disorder of a somewhat immature, aggressive, and rather dependent type was diagnosed. It was felt, however, that the possibility of temporal-lobe epilepsy could not be excluded. Unfortunately the patient did not keep an appointment for an electroencephalogram. Because of the combination of a personality disorder and his above-average height 12 the peripheral-blood leucocytes were karyotyped. Fifty cells were counted and analysed and twenty-six had 46 chromosomes with a normal sex-chromosome constitution, while nineteen 1. 2.
Price, W. H., Whatmore, P. B. Br. med. J. i, 533. Jacobs, P. A., Jacobs, A., Price, W. H., Court-Brown, W. M., Brittain, R. P., Whatmore, P. B. Ann. hum. Genet. 1968, 32, 339.
356 had 47 chromosomes and an XYY sex-chromosome constitution. The remaining five cells did not have a consistent analysis. The patient was reported as a 46, XY/47, XYY mosaic.
grateful to Dr. Patricia Jacobs for the report on the chromoconstitution, to Dr. W. H. Price for his advice, and to Mr. lain Paterson, psychiatric social worker, for information derived from the patient’s parents. M. B. MATTHEWS Western General Hospital, P. W. BROOKS. Edinburgh 4. We are
the variants of glucose-6-phosphate dehydrogenase), so that temporary nomenclature could be suggested. Laboratoire d’Enzymologie, Unité de Recherches Isotopiques de l’I.N.S.E.R.M., P. BOIVIN C. GALAND. Hôpital Beaujon, 92 Clichy, France.
a
some
ANTILYMPHOCYTE SERUM SIR,-Your annotation (July 27, p. 204) does less than justice to this important subject. The discovery that " antilymphocyteserum products may abolish the immunological opposition to grafts " is at least five years old, and even if you mean heterografts the discovery goes back to 19661 or even earlier. The effectiveness and advantages of IgG prepared from antilymphocyte serum (A.L.S.), instead of crude serum, have been stressed in many papers over the last few years from our own laboratory and elsewhere, including one in The Lancet,2 and a great deal of work has been done which throws light on the mode of action of A.L.S. in vivo and in vitro. We realise that space is limited but feel that you might have helped those of your readers who are unfamiliar with this work to gain a fuller and more balanced picture by citing one of two recent review articles. One such appeared last year3 and a fuller and more up-to-date review from the same author is to appear shortly.4 University Department Medical
of
Surgical Science,
School,
Edinburgh,
8.
7,X DA M. F. A WOODRUFF KEITH JAMES.
*** Not many months ago we had a leader5 on antilymphocyte serum, so our annotation simply aimed to pick up a few of the latest findings, of which there are a vast number.-ED. L.
VARIANTS OF ERYTHROCYTIC PYRUVATE KINASE SIR,-Professor Lohr and his colleagues 6have reported electrophoretic abnormalities of erythrocytic pyruvate kinase (P.K.) in hxmolysates of patients with congenital hxmolytic anxmia. In one of the cases described the Michaelis constant of the enzyme for phosphoenolpyruvate (P.E.P. Km) was high.’ We have studiedcertain characteristics of concentrated P.K., extracted from hxmolysates of ten normal subjects, seven patients with congenital hxmolytic non-spherocytic anxmia with homozygous deficiency in P.K., and six collateral relatives in whom the deficiency was present in a " heterozygous " state. Of the seven homozygous subjects, in three the P.E.P. Km was equal to that of normal subjects; in two, it was about a quarter of normal; and in two it was considerably increased, as in the case described by Lohr.’ In five of the six heterozygous subjects it was normal; in one it was moderately increased. The Michaelis constant for adenosine phosphate in three patients with congenital hxmolytic anaemia with P.K. deficiency was found to be equal to that of control subjects. Such findings confirm the existence of various types of molecular abnormalities in connection with P.K. deficiency. It therefore seems desirable that criteria should now be defined for characterisation of these various types (as has been done for 1. Monaco, A. P., Wood, M. L., Gray, J. G., Russell, P. S. J. Immun. 1966, 96, 229. 2. Anderson, N. F., James, K., Woodruff, M. F. A. Lancet, 1967, i, 1126. 3. James, K. Clin exp. Immun. 1967, 2, 615. 4. James, K. Progr. Surg. (in the press). 5. Lancet, 1967, ii, 661. 6. Blume, K. G., Löhr, G. W., Rüdiger, H. W., Schalhorn, A. ibid. 1968,
i, 529. 7. Löhr, G. W., Blume, K. G., Rüdiger, H. W., Sokal, G., Gulbis, E. ibid. p. 753. 8. Boivin, P., Galand, C. Nouv. Revue fr. Hémat. 1968, 8, 201.
ALKALINE-PHOSPHATASE-SECRETING TUMOUR OF LUNG
SIR,- There have been several reports of tumours not involving bone, with a raised serum-alkaline-phosphatase. 12 The importance of these observations lies in the fact that the neoitself may be the source of the alkaline phosphatase. It is therefore important to characterise the different types of tumour alkaline phosphatase in order to determine whether there is any characteristic which is typical of any particular type of tumour. I have lately had a patient whose carcinoma of the lung contained a high concentration of alkaline phosphatase and who also had a raised level of enzyme in the serum. The patient was a man, aged 73 years, who presented at the
plasm
Manchester Royal Infirmary with a 6-month history of weight loss, hxmoptysis, and dyspnoea. Chest X-ray suggested a neoplasm in the left upper-lobe bronchus. Investigations were limited in view of the patient’s age and physical state, but he was found to have a serumalkaline-phosphatase of 20 King-Armstong units on three occasions. He died a month after admission. Necropsy showed an infiltrating carcinoma approximately 4-0 cm. in diameter in the left upper-lobe bronchus. Secondary deposits were present in the left kidney, both adrenal glands, and the para-aortic, cervical, and right axillary lymphnodes, and a small solitary deposit was present in the left lobe of the liver. There were no detectable bone secondaries, and the bileduct was patent. Histologically the tumour was a moderately welldifferentiated squamous-cell carcinoma. Cryostat sections were cut from both theprimary tumour and the secondary deposits, and the sections were stained for alkaline phosphatase by the method of Burstone.3 Both stained strongly. Alkaline phosphatase was extracted from the tumour with butanol,4 and disc electrophoresis was carried out using 5-0% acrylamide gel in a Shandon electrophoresis tank. The enzyme extracted from the tumour was run in parallel with a sample of serum from a case of bone disease (Paget’s disease), and the gels were stained in the same "
"
medium as was used for the cryostat sections. The tumour extract showed two bands: a strong band moving slightly more slowly than the fastest band in the serum, and a weaker band nearer the origin. The effect of heat on the enzyme was determined by removing samples at intervals from tumour extract incubated at 56°C. The amount of enzyme left in each sample was determined by the department of biochemistry at Manchester Royal Infirmary. The results were as follows:
There was 66% inhibition at 10 minutes and 83% at 30 minutes, after which there was no further inhibition. Electrophoresis of a sample after heat inactivation showed that all the remaining activity was in the broad fast band. No inactivation of enzyme activity was seen in the presence of 0-005 M L-phenylalanine. The idea that a tumour may be the source of a raised serum-
alkaline-phosphatase has been put forward before, but so far there are few studies of the biochemical properties of the types of alkaline phosphatase found in tumours. Fishman et a1.2 reported a case of a patient with carcinoma of the lung with metastases in lymph-nodes and other organs in which both the tumour and serum showed an atypical alkaline phosphatase in that it contained a very heat-stable L-phenylalanine-sensitive enzyme which accounted for 50% of the total serum activity. The enzyme from the present tumour differs from this in that it was largely heat-labile and insensitive to L-phenylalanine. Considerably more work will need to be done before any pattern can be associated with any particular tumour type. Department of Pathology, University, Manchester 13.
The
W. R. TIMPERLEY.
1. Schlang, H. A., McHenry, L. E., Jr. N.Y. State J. Med. 1962, 62, 3811. 2. Fishman, W. H., Inglis, Norma I., Stolbach, L. L., Krant, M. J. Cancer Res. 1968, 28, 150. 3. Burstone, M. S. J. natn. Cancer Inst. 1958, 20, 601; J. Histochem. Cytochem. 1958, 6, 322. 4. Morton, R. K. Biochem. J. 1954, 57, 595.