CLOSTRIDIAL TOXIN IN FÆCES OF HEALTHY INFANTS
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319 in this study were chosen to give a wide range of incubation periods, intensity and distribution of neuronal and white- matter vacuolation, and neuritic plaque formation. Brains were removed from normal, scrapie-affected and s.F.v.-affected male C57BL mice and stored at -25 to —30°C for up to six months before assay. Scrapie mice were killed when at a moderately advanced clinical stage, but when they would not have died of the disease for at least a week. The incubation period for the s.F.v.-infected mice ranged from 5 to 10z days when the disease was at a moderate to very advanced stage; some of the mice were in a more advanced state than any of the scrapie mice. C.A.T. was determined by a radiochemical method7 on homogenates of forebrain, midbrain, cerebellum, and hind- brain. The decay in activity of C.A.T. on storage at -25 to - 30°C was 3-4% per annum. The activity of C.A.T. was similar in forebrain, midbrain, and hindbrain, whereas the cerebellum had about a fifth of the activity of these regions (see table). There was no reduction in brain c.A.T. activity in s.F.Y.-infected mice. In the scrapie mouse brains, C.A.T. activity was usually reduced, the effect being most apparent in the 79A strain. All four regions show reduced ac- tivities. Comparison of all the scrapie-infected brains with the normal control group revealed significant differences (P<0.05, Mann-Whitney U test) for all brain regions. When results for individual scrapie agents were tested against normal controls at least one brain region had significantly reduced C.A.T. acti- vity in every instance. The importance and specificity of this finding-in particular how it relates to the reduced C.A.T. acti- vity in Alzheimer’s disease-remain to be determined, but a biochemical lesion common to Alzheimer’s disease and an ani- mal disease known to be due to a transmissible agent is clearly worthy of further investigation. M.R.C. Demyelinating Diseases Unit, Newcastle General Hospital, Newcastle upon Tyne NE4 6BE J. R. MCDERMOTT Moredun Institute, Gilmerton, Edinburgh H. FRASER A.R.C. Animal Breeding Research Organisation, Edinburgh A. G. DICKINSON CLOSTRIDIAL TOXIN IN FÆCES OF HEALTHY INFANTS SIR,—Dr Larson and co-workers found Clostridium6 difficile toxin in the fxces of two healthy neonates. In our laboratory the cytotoxic effect, now known to be caused by C. difficile toxin, was noted some years ago, in fæces sent in for virological examination but this observation could not be explained at that time. We have examined all records of virolo- gical fsecal cultures in 1976 and 1977 with respect to the age of the patients and the frequency of a positive cytotoxic effect. The samples were from inpatients. Positive results were rare in patients above the age of one year, but not rare in infants: Most positive samples in infants came from patients with res- piratory disease, convulsions, meningoencephalitis, or congeni- tal malformations. Only 7 had diarrhoea. It is very improbable that all these patients had pseudomembranous colitis. A toxin titre could be estimated in the positive samples because the fxcal extracts were diluted and subcultured in a standard way. This titre, on human diploid cells (embryonic 7. Fonnum, F. J. Neurochem. 1975, 24, 407. 1. Larson, H. E., Price, A. B., Honour, P., Borriello, S. P. Lancet, 1978, i, 1063. lung fibroblasts), was usually around 1:100 and about 1:1000 at most. Although such titres are often found in later stages of pseudomembranous colitis the titre of a fresh sample in the acute phase of the disease was, in our hands, much higher (1:56 000). Fxcal extracts were inoculated on primary monkey kidney cells as well. These cells turned out to be 10-100 times less sensitive to the toxin than the human diploid cells. With most positive samples there was no toxic effect on the monkey kidney cells and, when there was the effect was only slight. The cytotoxic effect in three recently obtained toxic fmces from infants could be neutralised by C. sordellii antitoxin (kindly supplied by Wellcome Research Laboratories). In no case was pseudomembranous colitis present and in one patient (titre 1:1400) a normal intestine was found at necropsy (this patient died from an unrelated disease). In several of these "control" infants C. difficile toxin was present in .the faeces, although at titres lower than those found in the acute phase of pseudomembranous colitis; probably toxin was of no pathological significance in these patients. These findings accord with the fact that C. difficile has been isolated from about 10% of the stools of infants.2 In contrast, C. difficile is rarely identified in the faeces of older persons.3 The preponderance of C. difficile in infants might be due to the presence of this microorganism in the vagina of many women.4 Laboratorium voor Medische Microbiologie, Academisch Ziekenhuis, Universiteit van Amsterdam, Amsterdam, Netherlands P. J. G. M. RIETRA K. W. SLATERUS H. C. ZANEN Binnengasthuis, Universiteit van Amsterdam S. G. M. MEUWISSEN CHRONIC PHENOTHIAZINE THERAPY DOES NOT INCREASE SELLAR SIZE SIR,—Several conditions other than primary disease of the pituitary increase pituitary size.5-8 In pregnancy the pituitary can enlarge due to lactotroph hyperplasia,’ but sellar enlarge- ment and remodelling do not usually ensue, pregnancy being a short-term condition in this context. Primary hypothy- roidism6 and primary gonadal failure7 8 have been associated with sellar enlargement, visual-field abnormalities, and hypo- pituitarism. PHENOTHIAZINE DOSE, SERUM-PROLACTIN, AND SELLAR AREA IN TWENTY-ONE MALES BEFORE AND AFTER LONG-TERM PHENOTHIAZINE S Values as mean ± s.D. (and range). * Dose at time serum-prolactin drawn, in chlorpromazine equivalents. t Normal 12.0 ng/ml. ‡1 square inch = 6.45 cm2. Because phenothiazines stimulate prolactin secretion we measured serum-prolactin concentrations in twenty-one males on long-term phenothiazine therapy and calculated the lateral sellar surface area by planimetry. All patients had skull X-rays taken on the day of prolactin measurement, and also on a pre- 2. Snyder, M. L. J. infect. Dis. 1937, 60, 223. 3. George, W. L., Sutter, V. L., Finegold, S. M. ibid 1977, 136, 822. 4. Hafiz, S., McEntegart, M. G., Morton, R. S., Watkins, S. A. Lancet, 1975, ii, 420. 5. Goluboff, L. G., Ezrin, C. J. clin. Endocr. Metab. 1969, 29, 1533. 6. Vagenakis, A. G., Dole, K., Braverman, L. E. Ann. intern. Med. 1976, 85, 195. 7. Kelly. L. W. J. clin. Endocr. Metab. 1963, 23, 50. 8. Leiba, S., Landau, B., Ber, A. Acta endocr 1969, 60, 112.
Transcript of CLOSTRIDIAL TOXIN IN FÆCES OF HEALTHY INFANTS
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in this study were chosen to give a wide range of incubationperiods, intensity and distribution of neuronal and white-matter vacuolation, and neuritic plaque formation. Brains wereremoved from normal, scrapie-affected and s.F.v.-affected maleC57BL mice and stored at -25 to —30°C for up to six monthsbefore assay. Scrapie mice were killed when at a moderatelyadvanced clinical stage, but when they would not have diedof the disease for at least a week. The incubation period forthe s.F.v.-infected mice ranged from 5 to 10z days when thedisease was at a moderate to very advanced stage; some of themice were in a more advanced state than any of the scrapiemice. C.A.T. was determined by a radiochemical method7 onhomogenates of forebrain, midbrain, cerebellum, and hind-brain. The decay in activity of C.A.T. on storage at -25 to- 30°C was 3-4% per annum.The activity of C.A.T. was similar in forebrain, midbrain, and
hindbrain, whereas the cerebellum had about a fifth of theactivity of these regions (see table). There was no reduction inbrain c.A.T. activity in s.F.Y.-infected mice. In the scrapie mousebrains, C.A.T. activity was usually reduced, the effect being mostapparent in the 79A strain. All four regions show reduced ac-tivities. Comparison of all the scrapie-infected brains with thenormal control group revealed significant differences (P<0.05,Mann-Whitney U test) for all brain regions. When results forindividual scrapie agents were tested against normal controlsat least one brain region had significantly reduced C.A.T. acti-vity in every instance. The importance and specificity of thisfinding-in particular how it relates to the reduced C.A.T. acti-vity in Alzheimer’s disease-remain to be determined, but abiochemical lesion common to Alzheimer’s disease and an ani-mal disease known to be due to a transmissible agent is clearlyworthy of further investigation.M.R.C. Demyelinating Diseases Unit,Newcastle General Hospital,Newcastle upon Tyne NE4 6BE J. R. MCDERMOTTMoredun Institute,Gilmerton, Edinburgh H. FRASER
A.R.C. Animal Breeding Research Organisation,Edinburgh A. G. DICKINSON
CLOSTRIDIAL TOXIN IN FÆCES OF HEALTHYINFANTS
SIR,—Dr Larson and co-workers found Clostridium6difficile toxin in the fxces of two healthy neonates. In ourlaboratory the cytotoxic effect, now known to be caused by C.difficile toxin, was noted some years ago, in fæces sent in for
virological examination but this observation could not be
explained at that time. We have examined all records of virolo-gical fsecal cultures in 1976 and 1977 with respect to the ageof the patients and the frequency of a positive cytotoxic effect.The samples were from inpatients. Positive results were rare inpatients above the age of one year, but not rare in infants:
Most positive samples in infants came from patients with res-piratory disease, convulsions, meningoencephalitis, or congeni-tal malformations. Only 7 had diarrhoea. It is very improbablethat all these patients had pseudomembranous colitis.A toxin titre could be estimated in the positive samples
because the fxcal extracts were diluted and subcultured in astandard way. This titre, on human diploid cells (embryonic
7. Fonnum, F. J. Neurochem. 1975, 24, 407.1. Larson, H. E., Price, A. B., Honour, P., Borriello, S. P. Lancet, 1978, i,
1063.
lung fibroblasts), was usually around 1:100 and about 1:1000at most. Although such titres are often found in later stages ofpseudomembranous colitis the titre of a fresh sample in theacute phase of the disease was, in our hands, much higher(1:56 000). Fxcal extracts were inoculated on primary monkeykidney cells as well. These cells turned out to be 10-100 timesless sensitive to the toxin than the human diploid cells. Withmost positive samples there was no toxic effect on the monkeykidney cells and, when there was the effect was only slight.The cytotoxic effect in three recently obtained toxic fmces
from infants could be neutralised by C. sordellii antitoxin
(kindly supplied by Wellcome Research Laboratories). In nocase was pseudomembranous colitis present and in one patient(titre 1:1400) a normal intestine was found at necropsy (thispatient died from an unrelated disease).
In several of these "control" infants C. difficile toxin waspresent in .the faeces, although at titres lower than those foundin the acute phase of pseudomembranous colitis; probablytoxin was of no pathological significance in these patients.These findings accord with the fact that C. difficile has beenisolated from about 10% of the stools of infants.2 In contrast,C. difficile is rarely identified in the faeces of older persons.3The preponderance of C. difficile in infants might be due to thepresence of this microorganism in the vagina of many women.4
Laboratorium voor Medische Microbiologie,Academisch Ziekenhuis,Universiteit van Amsterdam,Amsterdam, Netherlands
P. J. G. M. RIETRAK. W. SLATERUSH. C. ZANEN
Binnengasthuis,Universiteit van Amsterdam S. G. M. MEUWISSEN
CHRONIC PHENOTHIAZINE THERAPY DOES NOTINCREASE SELLAR SIZE
SIR,—Several conditions other than primary disease of thepituitary increase pituitary size.5-8 In pregnancy the pituitarycan enlarge due to lactotroph hyperplasia,’ but sellar enlarge-ment and remodelling do not usually ensue, pregnancy beinga short-term condition in this context. Primary hypothy-roidism6 and primary gonadal failure7 8 have been associatedwith sellar enlargement, visual-field abnormalities, and hypo-pituitarism.
PHENOTHIAZINE DOSE, SERUM-PROLACTIN, AND SELLAR AREA INTWENTY-ONE MALES BEFORE AND AFTER LONG-TERM
PHENOTHIAZINE S
Values as mean ± s.D. (and range).* Dose at time serum-prolactin drawn, in chlorpromazine equivalents.t Normal 12.0 ng/ml.‡1 square inch = 6.45 cm2.
Because phenothiazines stimulate prolactin secretion wemeasured serum-prolactin concentrations in twenty-one maleson long-term phenothiazine therapy and calculated the lateralsellar surface area by planimetry. All patients had skull X-raystaken on the day of prolactin measurement, and also on a pre-
2. Snyder, M. L. J. infect. Dis. 1937, 60, 223.3. George, W. L., Sutter, V. L., Finegold, S. M. ibid 1977, 136, 822.4. Hafiz, S., McEntegart, M. G., Morton, R. S., Watkins, S. A. Lancet, 1975,
ii, 420.5. Goluboff, L. G., Ezrin, C. J. clin. Endocr. Metab. 1969, 29, 1533.6. Vagenakis, A. G., Dole, K., Braverman, L. E. Ann. intern. Med. 1976, 85,
195.7. Kelly. L. W. J. clin. Endocr. Metab. 1963, 23, 50.8. Leiba, S., Landau, B., Ber, A. Acta endocr 1969, 60, 112.
