584 whether grossly elevated or borderline, the kit alert. Plans can then be made for an antidote regimen, at the same time as more refined (and time-consuming) analyses are carried out in the laboratory. This approach can be very helpful to clinicians in district general hospitals and, probably, no less helpful in many teaching centres. We agree that ward staff should be taught how to use the kit by an experienced laboratory worker. With that proviso, we think it is wrong to say that the results will be substantially misleading and to suggest that there is any danger in handling the kit reagents.
higher levels, will sound
an
Poisons Unit,
Guy’s Hospital, London
from the subjects with ulcerative colitis were distributed differently from other groups, however, and this seemed to be due to a preponderance of subjects with total colonic disease having low M.P.O. activity. Disease activity, distribution in Crohn’s disease and drug therapy seemed to have no influence on results. Since m.p.o. is a haem enzyme, the possibility was considered that low leucocyte M.p.o. might be related to serumiron. In retrospect serum-iron levels were available in seventeen patients (Crohn’s disease and colitis) but only 4 low
B. WIDDOP R. GOULDING
NEUTROPHIL FUNCTION AND MYELOPEROXIDASE ACTIVITY IN INFLAMMATORY BOWEL DISEASE to read of the abnormality of neufunction in Crohn’s disease reported by Segal and Loewi.1 Investigation of this cell is certainly relevant since defective neutrophil function with impaired reduction of nitroblue tetrazolium (N.B.T.) characterises chronic granulomatous disease,2 in which the gastrointestinal features closely resemble those of Crohn’s disease.3 Our own studies of N.B.T. reduction in Crohn’s disease, like those of Segal and Loewi did not show any impaired N.B.T. reduction,4 in fact in most cases N.B.T. reduction was enhanced. Although this may simply be a non-specific marker of inflammatory activity it is of interest, in view of the isolation of viruses in this condition3 that analysis of our results by the technique of Feigin et al.6 showed a tendency for results in Crohn’s disease to cluster in the "viral" area of the nomogram. We have also investigated neutrophil myeloperoxidase M.p.o., E.C. 1.11.1.7) in patients with inflammatory bowel disease because this enzyme is closely involved in intracellular killing of phagocytosed bacteria.7 Sixteen patients with Crohn’s disease, and twenty with ulcerative colitis conforming to the usually accepted diagnostic criteria8 9 were investigated and compared with seventeen healthy controls of similar age and sex distribution. A further group of subjects who had undergone total proctocolectomy for ulcerative colitis between 1 and 18 years previously was also studied. The majority of this group (sixteen) were female and all were in good health at the time of the study. Leucocytes were separated from heparinised blood within 3 h of venepuncture by a modification of the dextran sedimentation technique of Lehrer. 10 Contaminating erythrocytes were removed by differential hypotonic lysis using 0.87% w/v ammonium chloride. After resuspending the cells in lOmmol/1 acetate buffer, pH 3.8, lysis was achieved by homogenising for 60 s in a glass homogeniser with a ’teflon’ pestle followed by freezing and thawing three times in solid carbon dioxide. Debris was removed by centrifuging at 800g for 20 min. Myeloperoxidase activity in the supernatant was assayed by the method of Klebanoff" using o-dianisidine as substrate. Protein concentration was determined by the method of Miller,12 and results, which were expressed as units/µg protein, are shown in the figure. There were no significant differences between controls and any patient group (Kruskal-Wallis test). The results
SIR,—We were interested
trophil
1. Segal, A. W., Loewi, G. Lancet, 1976, ii, 219. 2. Park, B. H., Holmes, B. M., Rodey, G. E., Good, R. A. ibid. 1969, i, 157. 3. Ament, M. E., Ochs, H. D. New Engl. J. Med. 1967, 288, 382. 4. Ward, M., Eastwood, M. A. Digestion, 1976, 14, 179. 5. Gitmck, G. L., Arthur, M. H., Shibata, I. Lancet, 1976, ii, 215. 6. Feigin, R. D., Shackelford, P. G. Choi, S. C., Flake, K. K., Franklin, F A., Eisenberg, C. S. J. Pediat. 1971, 78, 230. 7. Klebanoff, S. J. A. Rev. Med. 1971, 22, 39. 8. Lennard Jones, J. E., Lockhart, Mummery, H. E., Morson, B. C. Gastroenterology, 1968, 54, 1162. 9. Schachter, H., Kirsner, J. B. ibid. 1975, 68, 591. 10. Lehrer, R. I. J. clin Invest., 1971, 50, 2498. 11. Klebanoff, S J. Endocrinology, 1965, 76, 301.
Neutrophil
M.P.O.
activity
in controls and in patients with Crohn’. a group after colectomy for
disease, with ulcerative colitis and in ulcerative coUtis.
enzyme activities were associated with low serum-iron concentrations and no significant linear correlation could be found (r
= 0.13). Since the results in the post-colectomy group do not show the same distribution seen in the ulcerative-colitis patients, this is probably a reflection of the prevailing inflammatory activity rather than any predisposing factor. A phagocyte defect also seems an unlikely primary setiological factor in inflammatory bowel disease in view of the absence of any impaired resistance to infection. However, disorders of neutrophil function have been demonstrated in conditions of known viral" and bacterial14 aetiology, and such defects might well contribute to the persistence of inflammation in gut mucosa damaged by some other primary factor. The combination of a common extrinsic agent such as a virus and a variable defect in neutrophil function might perhaps account for some of the perplexing similarities and differences between ulcerative colitis and Crohn’s disease.15
Department of Pædiatric Biochemistry and Clinical
Chemistry, Royal Hospital for Sick Children, Edinburgh Gastrointestinal Unit, University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU
Department of Pædiatric Biochemistry and Clinical Chemistry, Royal Hospital for Sick Children, Edinburgh
M. RENZ M. WARD M. A. EASTWOOD
R. A. HARKNESS
12. Miller, G. L. Analyt. Chem. 1959, 31, 964. 13. Craft, A. W., Reid, M. M., Low, W. T. Br. med. J. 1976, i, 1570. 14. Ward, P. A., Goralnick, S., Bullock, W. E. J. Lab. clin. Med. 1976, 87, 1025 15. Hywel Jones, J., Lennard Jones, J. E., Morson, B. C., Chapman, M., Sackin, M. J., Sneath, P. H. A., Spicer, C. L., Card, W. I. Q. Jl. Med. 1973, 42, 715.
higher levels, will sound
an
Poisons Unit,
Guy’s Hospital, London
from the subjects with ulcerative colitis were distributed differently from other groups, however, and this seemed to be due to a preponderance of subjects with total colonic disease having low M.P.O. activity. Disease activity, distribution in Crohn’s disease and drug therapy seemed to have no influence on results. Since m.p.o. is a haem enzyme, the possibility was considered that low leucocyte M.p.o. might be related to serumiron. In retrospect serum-iron levels were available in seventeen patients (Crohn’s disease and colitis) but only 4 low
B. WIDDOP R. GOULDING
NEUTROPHIL FUNCTION AND MYELOPEROXIDASE ACTIVITY IN INFLAMMATORY BOWEL DISEASE to read of the abnormality of neufunction in Crohn’s disease reported by Segal and Loewi.1 Investigation of this cell is certainly relevant since defective neutrophil function with impaired reduction of nitroblue tetrazolium (N.B.T.) characterises chronic granulomatous disease,2 in which the gastrointestinal features closely resemble those of Crohn’s disease.3 Our own studies of N.B.T. reduction in Crohn’s disease, like those of Segal and Loewi did not show any impaired N.B.T. reduction,4 in fact in most cases N.B.T. reduction was enhanced. Although this may simply be a non-specific marker of inflammatory activity it is of interest, in view of the isolation of viruses in this condition3 that analysis of our results by the technique of Feigin et al.6 showed a tendency for results in Crohn’s disease to cluster in the "viral" area of the nomogram. We have also investigated neutrophil myeloperoxidase M.p.o., E.C. 1.11.1.7) in patients with inflammatory bowel disease because this enzyme is closely involved in intracellular killing of phagocytosed bacteria.7 Sixteen patients with Crohn’s disease, and twenty with ulcerative colitis conforming to the usually accepted diagnostic criteria8 9 were investigated and compared with seventeen healthy controls of similar age and sex distribution. A further group of subjects who had undergone total proctocolectomy for ulcerative colitis between 1 and 18 years previously was also studied. The majority of this group (sixteen) were female and all were in good health at the time of the study. Leucocytes were separated from heparinised blood within 3 h of venepuncture by a modification of the dextran sedimentation technique of Lehrer. 10 Contaminating erythrocytes were removed by differential hypotonic lysis using 0.87% w/v ammonium chloride. After resuspending the cells in lOmmol/1 acetate buffer, pH 3.8, lysis was achieved by homogenising for 60 s in a glass homogeniser with a ’teflon’ pestle followed by freezing and thawing three times in solid carbon dioxide. Debris was removed by centrifuging at 800g for 20 min. Myeloperoxidase activity in the supernatant was assayed by the method of Klebanoff" using o-dianisidine as substrate. Protein concentration was determined by the method of Miller,12 and results, which were expressed as units/µg protein, are shown in the figure. There were no significant differences between controls and any patient group (Kruskal-Wallis test). The results
SIR,—We were interested
trophil
1. Segal, A. W., Loewi, G. Lancet, 1976, ii, 219. 2. Park, B. H., Holmes, B. M., Rodey, G. E., Good, R. A. ibid. 1969, i, 157. 3. Ament, M. E., Ochs, H. D. New Engl. J. Med. 1967, 288, 382. 4. Ward, M., Eastwood, M. A. Digestion, 1976, 14, 179. 5. Gitmck, G. L., Arthur, M. H., Shibata, I. Lancet, 1976, ii, 215. 6. Feigin, R. D., Shackelford, P. G. Choi, S. C., Flake, K. K., Franklin, F A., Eisenberg, C. S. J. Pediat. 1971, 78, 230. 7. Klebanoff, S. J. A. Rev. Med. 1971, 22, 39. 8. Lennard Jones, J. E., Lockhart, Mummery, H. E., Morson, B. C. Gastroenterology, 1968, 54, 1162. 9. Schachter, H., Kirsner, J. B. ibid. 1975, 68, 591. 10. Lehrer, R. I. J. clin Invest., 1971, 50, 2498. 11. Klebanoff, S J. Endocrinology, 1965, 76, 301.
Neutrophil
M.P.O.
activity
in controls and in patients with Crohn’. a group after colectomy for
disease, with ulcerative colitis and in ulcerative coUtis.
enzyme activities were associated with low serum-iron concentrations and no significant linear correlation could be found (r
= 0.13). Since the results in the post-colectomy group do not show the same distribution seen in the ulcerative-colitis patients, this is probably a reflection of the prevailing inflammatory activity rather than any predisposing factor. A phagocyte defect also seems an unlikely primary setiological factor in inflammatory bowel disease in view of the absence of any impaired resistance to infection. However, disorders of neutrophil function have been demonstrated in conditions of known viral" and bacterial14 aetiology, and such defects might well contribute to the persistence of inflammation in gut mucosa damaged by some other primary factor. The combination of a common extrinsic agent such as a virus and a variable defect in neutrophil function might perhaps account for some of the perplexing similarities and differences between ulcerative colitis and Crohn’s disease.15
Department of Pædiatric Biochemistry and Clinical
Chemistry, Royal Hospital for Sick Children, Edinburgh Gastrointestinal Unit, University of Edinburgh, Western General Hospital, Edinburgh EH4 2XU
Department of Pædiatric Biochemistry and Clinical Chemistry, Royal Hospital for Sick Children, Edinburgh
M. RENZ M. WARD M. A. EASTWOOD
R. A. HARKNESS
12. Miller, G. L. Analyt. Chem. 1959, 31, 964. 13. Craft, A. W., Reid, M. M., Low, W. T. Br. med. J. 1976, i, 1570. 14. Ward, P. A., Goralnick, S., Bullock, W. E. J. Lab. clin. Med. 1976, 87, 1025 15. Hywel Jones, J., Lennard Jones, J. E., Morson, B. C., Chapman, M., Sackin, M. J., Sneath, P. H. A., Spicer, C. L., Card, W. I. Q. Jl. Med. 1973, 42, 715.
