695 IMMUNOFLUORESCENCE OF HUMAN REOVIRUS-LIKE AGENT OF INFANTILE DIARRHŒA
SIR,-Wyatt et al.’ described the successful cultivation ( human reovirus-like (R.v.L.) agent from stool filtrates in pr mary human embryonic kidney (H.E.K.) cell-culture. In Japan, R.V.L. agent is found in infantile diarrhoea accon panied by white stools (called hakuri in Japanese).2 The caus of the white stools is still unclear. We have tried to titral serum-antibody of infantile diarrhoea by indirect immum fluorescence (LF.), using primary H.E.K. cells infected wit antigen source. was prepared from white-diarrhoea sto( from an 11-month-old child admitted to the pacdiatric ward ( Nagoya University Hospital. Acute and convalescent sera wet collected from 7 other infants (aged 7-13 mo) with white-sto( diarrhrea. The stool filtrates of these 8 patients were examine by electron microscopy, and large quantities of R.V.L. ager were identified in all stools. 1.5 aliquots of the 2% stool fi trates were inoculated on primary H.E.K. monolayers in sma
R.V.L.
agent
as
A 2% stool filtrate3
initial choict of digoxin dose.
’Daily dose: first maintenance dose is given 6 h after loading dose. "Patients requiring reduced doses can be identified by routine -e measurement of serum-creatinine concentration and prediction of 10 creatinine clearance from nomogram of Siersbaek-Nielson et al.
tion of both loading2 6 and maintenance78 doses are necessary for greater degrees of impairment. The short interval between loading and maintenance dose allows the former to beBI small, so reducing the risk of early gastrointestinal symptoms. Department of Pharmacology and Therapeutics, Middlesex Hospital Medical School, London W1P 7PN
SYLVIA M. DOBBS
culture bottles. These infected cell-cultures were maintaine with Eagle M.E.M. at 37°C. Although inoculated monolaye; showed slight degenerative changes, clear cytopathic effect WI not observed. On the 14th day after inoculation the cultun were frozen and thawed once, and the cell-fluid mixture WI inoculated into other primary H.E.K. cell-cultures. At this tin infected cells were removed from the bottles with try] sin-E.D.T.A. and concentrated by centrifugation; and aceton fixed preparations were made for l.F. Convalescent serum froi 1 patient and F.I.T.c.-conjugated rabbit anti-human IgM we] used for staining. To check the successful passage of the agen Of. was done after every passage. Viral antigens, seen as fine granules in the cytoplasm, we: observed after each passage. After the fifth passage, the pe centage of I.F.-positive cells was estimated to be 3-4%.
Wyatt, R. G., Gill, V. W., Sereno, M. M., Kalica, A. R., Vankirk, D. H., Chanock, R. M., Kapikian, A. Z. Lancet, 1976, i, 98. 2. Konno, T., Suzuki, H., Ishida, N. ibid. 1975, i, 918. 3. Bishop, R. F., Davidson, G. P., Holmes, I. H., Ruck, B. J. ibid. 1974, i, 149. 4. Kapikian, A. Z., Kim, H. W., Wyatt, R. G., Rodriguez, W. J., Ross, S., Cline, W. L., Parrott, R. H., Chanock, R. M. Science, 1974, 185, 1049. 1.
ONDINE’S CURSE
SIR,-Dr Swift (Sept. 11,
p. 588) says he would welcome the case of Ondine’s curse he describes. At this point in our knowledge of sleep-linked respiratory periodicity may I suggest that the child should have a permanent tracheostomy with the valve open at night (and during the afternoon sleep if she has one) and closed by day. Quite a lot has been written on this subject-earlier this year, for example, Guilleminault et al. described sleep apnoea in comments on
eight children.11I If before a tracheostomy is done medical treatment can be tned I would suggest that clomipramine should be given in the erening (half the dose with the evening meal and half at bedtime This treatment
has been successful in adults with the
pickwickian syndrome 12 13 and has been tried with some success children with near-miss sudden-infant-death syndrome (unseems old enough for this treatment; in small babies with near-miss s.i.D.s. the problem is Tiore difficult because we do not know the effect of the drug un the maturing brain.
In
publishedl. Dr Swift’s patient
Laboratoire d’E.E G., G., Hôoital Broussais, Paris XIV, France
B. A. SCHWARTZ
6 Reuning, R.H., Sams, R. A., Notan, R. E. J. clin. Pharmac. 1973, 13, 127. 7 Jelif e, R. W., Brooker, G. Am. J. Med. 1974, 57, 63. 8 Dettli, L., Spring, P., Ryter, S. Acta Pharmac. Tox. 1971, 29, suppl. 3, 211. d, H. Cattell, M., Modell, W., Grainer, T., Guevara, R. J. Pharmac. exp. Ther 1950, 98, 337 10 Siersbaek-Nielson, K, Hansen, J. M., Kampmann, U., Kristensen, M. 9 Gol
Lancet, 1971, i, 1133. ault, C., Eldridge, F. L., Simmons, F. B., Dement, W. C. Pediatrics, 1976, 58, 23. 12 Schwartz, B A, Granelet-Eprinchard, M.-F. Revue E.E.G. Neurophysiol. 1974, 4, 79 13 Schwartz, B A, Rochemaure, J. Nouv Presse méd 1973, 2, 1520. 11 Guillemin
Fig. 1-IgG activities in sera of 7 patients.
SIR,-Wyatt et al.’ described the successful cultivation ( human reovirus-like (R.v.L.) agent from stool filtrates in pr mary human embryonic kidney (H.E.K.) cell-culture. In Japan, R.V.L. agent is found in infantile diarrhoea accon panied by white stools (called hakuri in Japanese).2 The caus of the white stools is still unclear. We have tried to titral serum-antibody of infantile diarrhoea by indirect immum fluorescence (LF.), using primary H.E.K. cells infected wit antigen source. was prepared from white-diarrhoea sto( from an 11-month-old child admitted to the pacdiatric ward ( Nagoya University Hospital. Acute and convalescent sera wet collected from 7 other infants (aged 7-13 mo) with white-sto( diarrhrea. The stool filtrates of these 8 patients were examine by electron microscopy, and large quantities of R.V.L. ager were identified in all stools. 1.5 aliquots of the 2% stool fi trates were inoculated on primary H.E.K. monolayers in sma
R.V.L.
agent
as
A 2% stool filtrate3
initial choict of digoxin dose.
’Daily dose: first maintenance dose is given 6 h after loading dose. "Patients requiring reduced doses can be identified by routine -e measurement of serum-creatinine concentration and prediction of 10 creatinine clearance from nomogram of Siersbaek-Nielson et al.
tion of both loading2 6 and maintenance78 doses are necessary for greater degrees of impairment. The short interval between loading and maintenance dose allows the former to beBI small, so reducing the risk of early gastrointestinal symptoms. Department of Pharmacology and Therapeutics, Middlesex Hospital Medical School, London W1P 7PN
SYLVIA M. DOBBS
culture bottles. These infected cell-cultures were maintaine with Eagle M.E.M. at 37°C. Although inoculated monolaye; showed slight degenerative changes, clear cytopathic effect WI not observed. On the 14th day after inoculation the cultun were frozen and thawed once, and the cell-fluid mixture WI inoculated into other primary H.E.K. cell-cultures. At this tin infected cells were removed from the bottles with try] sin-E.D.T.A. and concentrated by centrifugation; and aceton fixed preparations were made for l.F. Convalescent serum froi 1 patient and F.I.T.c.-conjugated rabbit anti-human IgM we] used for staining. To check the successful passage of the agen Of. was done after every passage. Viral antigens, seen as fine granules in the cytoplasm, we: observed after each passage. After the fifth passage, the pe centage of I.F.-positive cells was estimated to be 3-4%.
Wyatt, R. G., Gill, V. W., Sereno, M. M., Kalica, A. R., Vankirk, D. H., Chanock, R. M., Kapikian, A. Z. Lancet, 1976, i, 98. 2. Konno, T., Suzuki, H., Ishida, N. ibid. 1975, i, 918. 3. Bishop, R. F., Davidson, G. P., Holmes, I. H., Ruck, B. J. ibid. 1974, i, 149. 4. Kapikian, A. Z., Kim, H. W., Wyatt, R. G., Rodriguez, W. J., Ross, S., Cline, W. L., Parrott, R. H., Chanock, R. M. Science, 1974, 185, 1049. 1.
ONDINE’S CURSE
SIR,-Dr Swift (Sept. 11,
p. 588) says he would welcome the case of Ondine’s curse he describes. At this point in our knowledge of sleep-linked respiratory periodicity may I suggest that the child should have a permanent tracheostomy with the valve open at night (and during the afternoon sleep if she has one) and closed by day. Quite a lot has been written on this subject-earlier this year, for example, Guilleminault et al. described sleep apnoea in comments on
eight children.11I If before a tracheostomy is done medical treatment can be tned I would suggest that clomipramine should be given in the erening (half the dose with the evening meal and half at bedtime This treatment
has been successful in adults with the
pickwickian syndrome 12 13 and has been tried with some success children with near-miss sudden-infant-death syndrome (unseems old enough for this treatment; in small babies with near-miss s.i.D.s. the problem is Tiore difficult because we do not know the effect of the drug un the maturing brain.
In
publishedl. Dr Swift’s patient
Laboratoire d’E.E G., G., Hôoital Broussais, Paris XIV, France
B. A. SCHWARTZ
6 Reuning, R.H., Sams, R. A., Notan, R. E. J. clin. Pharmac. 1973, 13, 127. 7 Jelif e, R. W., Brooker, G. Am. J. Med. 1974, 57, 63. 8 Dettli, L., Spring, P., Ryter, S. Acta Pharmac. Tox. 1971, 29, suppl. 3, 211. d, H. Cattell, M., Modell, W., Grainer, T., Guevara, R. J. Pharmac. exp. Ther 1950, 98, 337 10 Siersbaek-Nielson, K, Hansen, J. M., Kampmann, U., Kristensen, M. 9 Gol
Lancet, 1971, i, 1133. ault, C., Eldridge, F. L., Simmons, F. B., Dement, W. C. Pediatrics, 1976, 58, 23. 12 Schwartz, B A, Granelet-Eprinchard, M.-F. Revue E.E.G. Neurophysiol. 1974, 4, 79 13 Schwartz, B A, Rochemaure, J. Nouv Presse méd 1973, 2, 1520. 11 Guillemin
Fig. 1-IgG activities in sera of 7 patients.
