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SUPPLY OF FACTOR VIII
213
donor, would allow
a donor to give more plasma for factor-vm separation. 2. Storage of the precipitate in the frozen state is usually feasible only at source, and the product can be supplied on demand only for immediate use. A small-scale trial has shown that pooling and freezedrying of cryoglobulin would not only facilitate transport and storage but also maintain stability for longer periods.
Department of Pathology, Shenley Hospital, Hertfordshire.
F. NOUR-ELDIN.
PHYTOHÆMAGGLUTININ AND CANCER SiR,—I have read with interest the letter of Dr. Rubio and Dr. Unsgaard,l following mine2 in which I reported the Fig. 1—Pancreas of normal subject stained with (a) T.A.F./safranin and (b) A.F./safranin, showing identical staining of P-cells by the two methods. (Reduced to about a half from x 400.)
Fig. 2-Pancreas of diabetic subject showing staining of insular &bgr;-cells (arrowed) by A.F. (b), not revealed by T.A.F. (a). (Reduced to about a half from x 400.) obtained with T.A.F. and A.F. staining are identical (fig. 1). In the islets of diabetic subjects, on the contrary, the A.F. method, besides staining the same &bgr;-cells which were coloured with T.A.F., stains other cells of P-type as well (fig. 2). Pretreatment with potassium cyanide does not modify the results of the A.F. reaction, whereas it makes the T.A.F. reaction negative: this supports the hypothesis that for positive A.F. staining, only the A-insulin chain, containing the easily oxidisable disulphide groups, is necessary, while a positive T.A.F. reaction depends on the presence of both A and B chains, as well as on the specific 5 arrangement of some basic aminoacids in the B chain. These findings suggest that two types (at least) of -cells, which possibly secrete different insulins, can be found in the pancreatic islets of diabetic subjects. In one type, this hormone should have a structure partly similar to the normal insulin (positive A.F. reaction), and partly different from it (negative T.A.F. reaction); while, in the other type, the insulin reacts identically to both stains. ANTONIO M. MANCINI Institute of Morbid Anatomy, GLAUCO FRIZZERA of University Bologna, VECCHI. ADALBERTO Italy.
rationale of tumour treatment by phytohamiagglutinin (P.H.A.). Since the method used has not yet been published I should like to report it here in brief. 40 female Swiss BIH (Law) inbred-strain mice and 60 Swiss mice from a closed non-isogenic colony were used. Each animal received an intraperitoneal injection of 106 ascites tumour cells. The mice were divided into groups of 10 for the different treatment schedules, and in each case a group of mice served as control. 50 mg. P.H.A. (Burroughs Wellcome, London) was diluted in 5 ml. distilled water and 0.1 ml. was injected intravenously into the tail vein of each mouse. 7 days after tumour transplantation the peritoneal cavity of each mouse was opened, irrigated with saline, and the washings were collected. The volume of packed tumour cells in the washings was determined in calibrated centrifuge tubes. To test the significance of the difference between treated and control groups the Mann Whitney statistic was used, and was found to be much lower than expected in each experiment. Combining all these statistics into an overall test by the method of van Elteren we find z=—4-28, p< 0-0001, thus indicating that the tumour volume was significantly lower in the treated group. The lowest volume of tumour cells was obtained when P.H.A. was injected 72 hours before tumour transplantation. However, less tumour cells were also found when P.H.A. was given 24 hours before, on the day of, and 48 hours after, tumour
SUPPLY OF FACTOR VIII SIR,-Concentrates of antihaemophilic factor vm prepared by the cryoglobulin methodovercome many of the obstacles limiting the supply of human material. Nevertheless, there are two problems, solution of which in the following manner may gleady ease the situation: 1. Ltmited number of donations.-Although the cryoglobulin method C0:;serves the red
cells, the in-vivo survival of which does not seem affected, the donors are usually bled twice yearly only. The use =.‘ phsmapheresis, by permitting the return of the red cells to the 1 Pool, J. G., Hershgold, E. J., Pappenhagen, A. R. Nature, Lond. 1964, 203, 312; Pool, J. G., Shannon, A. E. Fedn Proc. Fedn Am. Socs exp. Biol. 1965, 24, 512.
transplantation. In further experiments the value of P.H.A. treatment in doses of 0-05 ml., 0-2 ml., and 0-3 ml. was investigated. It was found that the best results were obtained with 0-1ml. Varying the time of the administration it was seen that P.H.A. has no protective effect when it is administered 120 hours or more before tumour transplantation. Thus it seems that the action of P.H.A. is a very short-term one. This may explain why Dr. Rubio and Dr. Unsgaard did not obtain any protective effect from P.H.A. treatment. Department
of
Oncology,
Hadassah University Hospital,
Jerusalem.
E. ROBINSON.
VINCA ALKALOIDS AND SALIVARY-GLAND PAIN SiR,-Three patients receiving vincristine for acute leukxmia and two patients on vinblastine for Hodgkin’s disease being treated in this unit have had discomfort amounting to pain behind the angle of the jaw before the first bite of a meal which they are expecting to enjoy. This occurs on either the 2nd or 3rd day after the administration of either of these drugs, and may continue to occur sporadically for a day or two. The discomfort lasts for a minute or so, and is described as a stinging or soreness beneath the ears. In one patient it was of sufficient concern for a dental opinion to be sought and for the mandible to be X-rayed. I do not think that this effect has been previously described; in itself it might be considered neither of interest nor of any use -in fact I believe that it is a normal experience for some people. However, I have further noted that three of the patients became severely constipated 4 or 5 days after the drug 1.
2.
Rubio, C. A., Unsgaard, B. Lancet, 1966, ii, 1191. Robinson, E. ibid. p. 753.
donor, would allow
a donor to give more plasma for factor-vm separation. 2. Storage of the precipitate in the frozen state is usually feasible only at source, and the product can be supplied on demand only for immediate use. A small-scale trial has shown that pooling and freezedrying of cryoglobulin would not only facilitate transport and storage but also maintain stability for longer periods.
Department of Pathology, Shenley Hospital, Hertfordshire.
F. NOUR-ELDIN.
PHYTOHÆMAGGLUTININ AND CANCER SiR,—I have read with interest the letter of Dr. Rubio and Dr. Unsgaard,l following mine2 in which I reported the Fig. 1—Pancreas of normal subject stained with (a) T.A.F./safranin and (b) A.F./safranin, showing identical staining of P-cells by the two methods. (Reduced to about a half from x 400.)
Fig. 2-Pancreas of diabetic subject showing staining of insular &bgr;-cells (arrowed) by A.F. (b), not revealed by T.A.F. (a). (Reduced to about a half from x 400.) obtained with T.A.F. and A.F. staining are identical (fig. 1). In the islets of diabetic subjects, on the contrary, the A.F. method, besides staining the same &bgr;-cells which were coloured with T.A.F., stains other cells of P-type as well (fig. 2). Pretreatment with potassium cyanide does not modify the results of the A.F. reaction, whereas it makes the T.A.F. reaction negative: this supports the hypothesis that for positive A.F. staining, only the A-insulin chain, containing the easily oxidisable disulphide groups, is necessary, while a positive T.A.F. reaction depends on the presence of both A and B chains, as well as on the specific 5 arrangement of some basic aminoacids in the B chain. These findings suggest that two types (at least) of -cells, which possibly secrete different insulins, can be found in the pancreatic islets of diabetic subjects. In one type, this hormone should have a structure partly similar to the normal insulin (positive A.F. reaction), and partly different from it (negative T.A.F. reaction); while, in the other type, the insulin reacts identically to both stains. ANTONIO M. MANCINI Institute of Morbid Anatomy, GLAUCO FRIZZERA of University Bologna, VECCHI. ADALBERTO Italy.
rationale of tumour treatment by phytohamiagglutinin (P.H.A.). Since the method used has not yet been published I should like to report it here in brief. 40 female Swiss BIH (Law) inbred-strain mice and 60 Swiss mice from a closed non-isogenic colony were used. Each animal received an intraperitoneal injection of 106 ascites tumour cells. The mice were divided into groups of 10 for the different treatment schedules, and in each case a group of mice served as control. 50 mg. P.H.A. (Burroughs Wellcome, London) was diluted in 5 ml. distilled water and 0.1 ml. was injected intravenously into the tail vein of each mouse. 7 days after tumour transplantation the peritoneal cavity of each mouse was opened, irrigated with saline, and the washings were collected. The volume of packed tumour cells in the washings was determined in calibrated centrifuge tubes. To test the significance of the difference between treated and control groups the Mann Whitney statistic was used, and was found to be much lower than expected in each experiment. Combining all these statistics into an overall test by the method of van Elteren we find z=—4-28, p< 0-0001, thus indicating that the tumour volume was significantly lower in the treated group. The lowest volume of tumour cells was obtained when P.H.A. was injected 72 hours before tumour transplantation. However, less tumour cells were also found when P.H.A. was given 24 hours before, on the day of, and 48 hours after, tumour
SUPPLY OF FACTOR VIII SIR,-Concentrates of antihaemophilic factor vm prepared by the cryoglobulin methodovercome many of the obstacles limiting the supply of human material. Nevertheless, there are two problems, solution of which in the following manner may gleady ease the situation: 1. Ltmited number of donations.-Although the cryoglobulin method C0:;serves the red
cells, the in-vivo survival of which does not seem affected, the donors are usually bled twice yearly only. The use =.‘ phsmapheresis, by permitting the return of the red cells to the 1 Pool, J. G., Hershgold, E. J., Pappenhagen, A. R. Nature, Lond. 1964, 203, 312; Pool, J. G., Shannon, A. E. Fedn Proc. Fedn Am. Socs exp. Biol. 1965, 24, 512.
transplantation. In further experiments the value of P.H.A. treatment in doses of 0-05 ml., 0-2 ml., and 0-3 ml. was investigated. It was found that the best results were obtained with 0-1ml. Varying the time of the administration it was seen that P.H.A. has no protective effect when it is administered 120 hours or more before tumour transplantation. Thus it seems that the action of P.H.A. is a very short-term one. This may explain why Dr. Rubio and Dr. Unsgaard did not obtain any protective effect from P.H.A. treatment. Department
of
Oncology,
Hadassah University Hospital,
Jerusalem.
E. ROBINSON.
VINCA ALKALOIDS AND SALIVARY-GLAND PAIN SiR,-Three patients receiving vincristine for acute leukxmia and two patients on vinblastine for Hodgkin’s disease being treated in this unit have had discomfort amounting to pain behind the angle of the jaw before the first bite of a meal which they are expecting to enjoy. This occurs on either the 2nd or 3rd day after the administration of either of these drugs, and may continue to occur sporadically for a day or two. The discomfort lasts for a minute or so, and is described as a stinging or soreness beneath the ears. In one patient it was of sufficient concern for a dental opinion to be sought and for the mandible to be X-rayed. I do not think that this effect has been previously described; in itself it might be considered neither of interest nor of any use -in fact I believe that it is a normal experience for some people. However, I have further noted that three of the patients became severely constipated 4 or 5 days after the drug 1.
2.
Rubio, C. A., Unsgaard, B. Lancet, 1966, ii, 1191. Robinson, E. ibid. p. 753.