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DIPHTHERIA TOXIN OF HIGH VALUE.
402 are
formed
as
the result of breaking down of blood of cellular disintegration-i.e.,
Thus, out of 27 cases, in which disseminated sclerosis only accounted for five of the series, 20 conditions
protein by products
other than disseminated were positive for the spherule enzymes. To test this possibility, a normal, fresh brain was and two negative. stripped of its meninges, the grey and white matter CONCLUSIONS. separated as far as possible from blood-vessels, &c., (1) The spherule described by Chevassut is not and ground up with sharp sand. The resultant to spinal fluids of disseminated sclerosis, but specific filtered filters fine Berkefeld until mixture was through in the fluids from many other conditions. can be found it was optically clear. This extract was then used in with the published findings of Lepine, This agrees a broth and serum mixture in the place of the cerebroand Tronconi. Mollaret, (2) Many of the essential spinal fluid. In 24 to 48 hours an optical appearance that of the need for immediate conditions, especially and of filaments spherules, granules, consisting of the spinal fluid to the broth-serum indistinguishable from that obtained in disseminated transference laid down in the original paper appear to mixture, sclerosis was seen. be open to considerable elasticity on further investigaTABLE OF RESULTS. tion. (3) We have not been able to obtain agar slipCases. cultures, but this has been accomplished by Tronconi Cerebral thrombosis 1 Spherules present. 3 tumour .. Spherules present in all. in disseminated sclerosis and in other lesions of the 1 abseess .... nervous system. Spherules present. (4) We do not consider the spherule No spherules observed Spinal curvature 1 to be a virus, but a flocculation or precipitation after 5 days. sclerosis phenomenon produced by the interaction of the G.P.I. .. .. .. 4 . . . ; Spherules present in a]].I spinal fluid and blood-serum. (5) It has not been .. .. .. 1 Tabes 1 Spastic paraplegia possible to study a sufficient number of normal 1 Parkinson’s Disease in order to state whether this phenomenon fluids Subacute combined indicates an abnormal fluid or whether it occurs with degeneration of cord 1 Amyotrophic Spherules present. any fluid. sclerosis.. 1 Paralysis agitans 1 Epilepsy 1 In conclusion, it is desired to thank the physicians Amentia .. 1 of the Westminster Hospital and of the Springfield 1 Sciatica 1 .... Urseimo fits Mental Hospital, Tooting, for their assistance in Tuberculous meningitis.. 2 Spherules present in one fluids for these ....
....
"
....
"
....
Disseminated
5
....
..
..
..
I
}
..)
....
...
....
lateral
.
....
...
....
....
...
..
supplying
....
and absent in the other.
case
investigations.
I
The Table is a list of 27 cerebro-spinal fluids treated under the same conditions. The conditions were that each fluid was exposed on the bench for 48 hours before being transferred to the serum-broth mixtureI! (Hartley’s broth, pH 7’6). This mixture was passed through a small Berkefeld filter and A control broth-serum tube was similarly passed I through a filter and incubated. Examinations of control and test cultures .were made at intervals of 24 hours up to five days. i
REFERENCES.
1. Chevassut, K. : THE LANCET, 1930, i., 552. 2. Purves-Stewart, J. : Ibid., p. 560. 3. Smith, H. and Ransome, A. C. : Ibid., 1930, ii., 901. 4. Carmichael, E. A. : Proc. Roy. Soc. Med., 1931, xxiv., 591 ; see THE LANCET, 1931, i., 134. 5. Lépine, P., and Mollaret, P. : Bull. de l’Acad. de Méd., 1931,
then incubated.i 6.
i
cv., 512.
Tronconi, V. : Boll. della Soc. Medico-Chirurg. di Pavia, 1931,
ix., Fasc. 4. 7. Hicks, J. A. B., and Hocking, F. D. M.: THE LANCET, 1930, i., 612. 8. Hicks and Hocking: Ibid., 1931, i., 553 ; and Brit. Med. Jour., 1931, i., 425. 9. Welch, F. : Jour. Roy. Microscop. Soc., 1926, xvi., 262.
CLINICAL AND LABORATORY NOTES ! who
obtained the following results: M.L.D., 0-00038 c.cm. ; Lo., 0-022 c.cm. ; L+, 0-026 c.cm. I hope shortly to publish full details of the method of preparation of this medium and of others which have been found to give high-value toxins.
DIPHTHERIA TOXIN OF HIGH VALUE.
!
BY C. G. POPE, B.SC. BRISTOL, BIOCHEMIST,
WELLCOME PHYSIOLOGICAL RESEARCH
LABORATORIES.
I wish to express thanks to Dr. Hartley for the FOR a number of years we have steadily increased tests which he carried out on this toxin, and to Dr. the strength of the diphtheria toxin produced at the R. G. White for information regarding the methods Wellcome Physiological Research Laboratories, and used for preparing diphtheria toxin at the Belmont I feel it would be of interest to place on record the Laboratories. details of one batch of toxin of unusually high value. Working with tryptic digest medium, we have ANTITOXIN OF EXCEPTIONAL POTENCY. frequently obtained toxins with an Lf. value of BY A. T. GLENNY, B.SC. LOND., 40 units per c.cm.* Recently a medium prepared from horse-muscle infusion and proteose peptone, containSEROLOGIST, WELLCOME PHYSIOLOGICAL RESEARCH LABORATORIES. ing sodium acetate, and sterilised by filtration through a Berkefeld candle-both important pointsgave toxin with an Lf. value of 50 units per c.cm. IT is of interest to other workers to place on record This toxin was examined at the National Institute for instances of antitoxin of exceptional potency. Among Medical Research by Mr. Percival Hartley, D.Sc., a group of 52 horses recently immunised with diphtheria toxin containing an average of 25 Lf. units * The abbreviation Lf. is a descendant of Ehrlich’s Lo = Limes null—i.e., the dose of diphtheria antitoxin which, when mixed per c.cm., prepared by Mr. C. G. Pope, seven produced with one test dose of toxin, and injected into a guinea-pig, serum containing from 2000 to 2500 units of antitoxin will prevent the appearance of any local oedema ; similarly L + is the amount of antitoxin which just fails to prevent the death per c.cm., three between 2500 and 3000, and one of an animal. Lf. is the amount of toxin which (f) flocculates contained 3000 units per c.cm. As this is the strongest with one unit of antitoxin when mixed in the Ramon method. unconcentrated antitoxin yet produced at the —ED. L. .
formed
as
the result of breaking down of blood of cellular disintegration-i.e.,
Thus, out of 27 cases, in which disseminated sclerosis only accounted for five of the series, 20 conditions
protein by products
other than disseminated were positive for the spherule enzymes. To test this possibility, a normal, fresh brain was and two negative. stripped of its meninges, the grey and white matter CONCLUSIONS. separated as far as possible from blood-vessels, &c., (1) The spherule described by Chevassut is not and ground up with sharp sand. The resultant to spinal fluids of disseminated sclerosis, but specific filtered filters fine Berkefeld until mixture was through in the fluids from many other conditions. can be found it was optically clear. This extract was then used in with the published findings of Lepine, This agrees a broth and serum mixture in the place of the cerebroand Tronconi. Mollaret, (2) Many of the essential spinal fluid. In 24 to 48 hours an optical appearance that of the need for immediate conditions, especially and of filaments spherules, granules, consisting of the spinal fluid to the broth-serum indistinguishable from that obtained in disseminated transference laid down in the original paper appear to mixture, sclerosis was seen. be open to considerable elasticity on further investigaTABLE OF RESULTS. tion. (3) We have not been able to obtain agar slipCases. cultures, but this has been accomplished by Tronconi Cerebral thrombosis 1 Spherules present. 3 tumour .. Spherules present in all. in disseminated sclerosis and in other lesions of the 1 abseess .... nervous system. Spherules present. (4) We do not consider the spherule No spherules observed Spinal curvature 1 to be a virus, but a flocculation or precipitation after 5 days. sclerosis phenomenon produced by the interaction of the G.P.I. .. .. .. 4 . . . ; Spherules present in a]].I spinal fluid and blood-serum. (5) It has not been .. .. .. 1 Tabes 1 Spastic paraplegia possible to study a sufficient number of normal 1 Parkinson’s Disease in order to state whether this phenomenon fluids Subacute combined indicates an abnormal fluid or whether it occurs with degeneration of cord 1 Amyotrophic Spherules present. any fluid. sclerosis.. 1 Paralysis agitans 1 Epilepsy 1 In conclusion, it is desired to thank the physicians Amentia .. 1 of the Westminster Hospital and of the Springfield 1 Sciatica 1 .... Urseimo fits Mental Hospital, Tooting, for their assistance in Tuberculous meningitis.. 2 Spherules present in one fluids for these ....
....
"
....
"
....
Disseminated
5
....
..
..
..
I
}
..)
....
...
....
lateral
.
....
...
....
....
...
..
supplying
....
and absent in the other.
case
investigations.
I
The Table is a list of 27 cerebro-spinal fluids treated under the same conditions. The conditions were that each fluid was exposed on the bench for 48 hours before being transferred to the serum-broth mixtureI! (Hartley’s broth, pH 7’6). This mixture was passed through a small Berkefeld filter and A control broth-serum tube was similarly passed I through a filter and incubated. Examinations of control and test cultures .were made at intervals of 24 hours up to five days. i
REFERENCES.
1. Chevassut, K. : THE LANCET, 1930, i., 552. 2. Purves-Stewart, J. : Ibid., p. 560. 3. Smith, H. and Ransome, A. C. : Ibid., 1930, ii., 901. 4. Carmichael, E. A. : Proc. Roy. Soc. Med., 1931, xxiv., 591 ; see THE LANCET, 1931, i., 134. 5. Lépine, P., and Mollaret, P. : Bull. de l’Acad. de Méd., 1931,
then incubated.i 6.
i
cv., 512.
Tronconi, V. : Boll. della Soc. Medico-Chirurg. di Pavia, 1931,
ix., Fasc. 4. 7. Hicks, J. A. B., and Hocking, F. D. M.: THE LANCET, 1930, i., 612. 8. Hicks and Hocking: Ibid., 1931, i., 553 ; and Brit. Med. Jour., 1931, i., 425. 9. Welch, F. : Jour. Roy. Microscop. Soc., 1926, xvi., 262.
CLINICAL AND LABORATORY NOTES ! who
obtained the following results: M.L.D., 0-00038 c.cm. ; Lo., 0-022 c.cm. ; L+, 0-026 c.cm. I hope shortly to publish full details of the method of preparation of this medium and of others which have been found to give high-value toxins.
DIPHTHERIA TOXIN OF HIGH VALUE.
!
BY C. G. POPE, B.SC. BRISTOL, BIOCHEMIST,
WELLCOME PHYSIOLOGICAL RESEARCH
LABORATORIES.
I wish to express thanks to Dr. Hartley for the FOR a number of years we have steadily increased tests which he carried out on this toxin, and to Dr. the strength of the diphtheria toxin produced at the R. G. White for information regarding the methods Wellcome Physiological Research Laboratories, and used for preparing diphtheria toxin at the Belmont I feel it would be of interest to place on record the Laboratories. details of one batch of toxin of unusually high value. Working with tryptic digest medium, we have ANTITOXIN OF EXCEPTIONAL POTENCY. frequently obtained toxins with an Lf. value of BY A. T. GLENNY, B.SC. LOND., 40 units per c.cm.* Recently a medium prepared from horse-muscle infusion and proteose peptone, containSEROLOGIST, WELLCOME PHYSIOLOGICAL RESEARCH LABORATORIES. ing sodium acetate, and sterilised by filtration through a Berkefeld candle-both important pointsgave toxin with an Lf. value of 50 units per c.cm. IT is of interest to other workers to place on record This toxin was examined at the National Institute for instances of antitoxin of exceptional potency. Among Medical Research by Mr. Percival Hartley, D.Sc., a group of 52 horses recently immunised with diphtheria toxin containing an average of 25 Lf. units * The abbreviation Lf. is a descendant of Ehrlich’s Lo = Limes null—i.e., the dose of diphtheria antitoxin which, when mixed per c.cm., prepared by Mr. C. G. Pope, seven produced with one test dose of toxin, and injected into a guinea-pig, serum containing from 2000 to 2500 units of antitoxin will prevent the appearance of any local oedema ; similarly L + is the amount of antitoxin which just fails to prevent the death per c.cm., three between 2500 and 3000, and one of an animal. Lf. is the amount of toxin which (f) flocculates contained 3000 units per c.cm. As this is the strongest with one unit of antitoxin when mixed in the Ramon method. unconcentrated antitoxin yet produced at the —ED. L. .