1382 withSinemet ’ they occurred in 16 and were marked in 4. Like Professor Barbeau, we have found it difficult to correct these levodopa-related symptoms. Lowering the daily dosage or individual doses, although it attenuates the dystonic movements, is accompanied by re-emergence of more Parkinson’s-disease symptoms. However, in some found that administration of levodopa in we have patients doses at short intervals, to lessen fluctuations of plasmareduces the dyskinesias which occur dopa concentrations, " " off " effect. at the time of the on " or

treatment

Brookhaven National Laboratory, Associated Universities, Inc.,

Upton, Long Island, New York 11973, U.S.A.

EDUARDO S. TOLOSA WILLIAM E. MARTIN HAROLD P. COHEN.

C3 IN CYSTIC FIBROSIS SiR,-Conover and his colleagues reported elevated C3 levels in patients with cystic fibrosis (c.F.).1 In another study Hann and his colleagues reported raised C3 levels in 28 of 51 patients with c.F. and in 7 of 24 controls.2 However, we believe that serum-levels of individual complement components in children cannot be meaningfully interpreted unless they are compared with normal values obtained from age-matched controls. In a study of 163 healthy infants and children we found a statistically significant correlation of C3 levels with age, but not with

in the appropriate age-group (see figure). The projected high and low values using 95% confidence limits are included. Only 4 of the 23 c.F. patients have raised C3 levels; however, these 4 patients could not be distinguished from the other c.F. patients by clinical evaluation. 2. In groups 5, 6, and 7 the mean C3 level for the c.F. patients in each group was compared with the mean C3 level for all the normal subjects included in the same group. There was no significant difference between the means in any of these groups (p > 0-3 by Student’s t test for each group).

Also there was no apparent correlation of the C3 level with the sex or the Shwachman score of the c.F. patients. In contrast to previous reports, we did not find raised C3 levels in the majority of our c.F. patients when the values were compared with projected group mean values for the appropriate age, and we did not find a significant elevation of the mean C3 level for the c.F. patients in any of the age-groups tested. We should like to emphasise the importance of establishing normal values in any laboratory that measures complement profiles in childhood diseases. Department of Pediatrics, Children’s Hospital of

Philadelphia, University of Pennsylvania,

Philadelphia, Pennsylvania 19104, U.S.A.

THOMAS F. SCANLIN, JR. MICHAEL E. NORMAN MARY LORETTA ROSENLUND.

UNSATURATED FATTY ACIDS AND TRANSPLANTATION

C3 levels in children with C.F. and

age-matched controls.

race, and we also found a wide range of values within any given age-group.33 Therefore C3 levels were measured in 23 children with C.F., and the results were compared with the findings in normal age-matched controls. Samples of venous blood were allowed to clot for 2 hours at 4°C. Sera were then separated and aliquots were snap-frozen and stored at —70°C. C3 levels were determined in duplicate within two weeks of storage, using the technique of Mancini et awl.4 Immunoplates were bought from Hyland, Inc. (Division of Travenol Laboratories, Inc.,

sex or

Costa Mesa, California). The data

were

analysed in the following ways

:

SiR,—There is evidence that free polyunsaturated fatty acids suppress cellular immunity in vitro.1-3 However, experiments designed to study their effect in vivo must recognise their toxicity. Ring et al. reported that the mean survival-time of grafts from black inbred BD-6 rats onto white male Sprague-Dawley rats increased when the latter were treated with linoleic acid (L.A.). L.A. was toxic when given intraperitoneally (44% of the animals died within 5-10 days), and some orally treated animals had diarrhoea. Since the control group " remained entirely untreated" (i.e., free from treatment stress), it was difficult to determine how much of the observed immunological suppression was caused by a non-specific toxic effect and how much depended upon the specific suppression of the immune response. Ring et al. concluded that " only oral administration of unsaturated fatty acids can be recommended ". Subcutaneous injection of L.A. (a less stressful method of administration) prolonged the survival of tails grafted from female A mice onto female CBA mice, the controls being injected with saline.b Experiments have been carried out to assess the nonspecific stress effect operating in the above studies. 4 out of 5 female CBA mice died after receiving tail grafts from female A mice and daily subcutaneous injections of 100 1. L.A. (Sigma Chemicals). All 13 grafted animals died after daily subcutaneous injection of 100 jjd. oleic acid. Graft survival was prolonged in the single mouse which survived L.A. treatment (16 days as compared with 11-2±0-75 for a control group). In further experiments the effect ofNaudicelle’ oil (Bio-Oils Research, Nantwich), containing triglyceride esters of linoleic acid (70%) and y-linoleic acid (7%), has been tested. Although this material was not toxic

1. The C3 values for the c.F. patients were plotted against the projected group mean C3 values for 95 % of the general population

there was no difference in the mean survival-time of these tail grafts when 100 z.1. of oil was given subcutaneously

1. Conover, J. H., Conod, E. J., Hirschorn, K. Lancet, 1973, ii, 1501. 2. Hann, S., Holsclaw, D. S., Shin, H. S. ibid. 1974, ii, 520. 3. Norman, M. E., Gall, E. P., Taylor, A., Laster, L., Nilsson, U. R. J. Pediat. (in the press). 4. Mancini, G., Carbonara, H. O., Heremans, J. F. Immunochemistry,

1. Mertin, J., Shenton, B. K., Field, E. J. Br. med. J. 1973, ii, 777. 2. Field, E. J., Shenton, B. K., Joyce, G. ibid. 1974, i, 412. 3. Mertin, J., Hughes, D., Shenton, B. K., Dickinson, J. P. Klin. Wschr. 1974, 52, 248. 4. Ring, J., Seifert, J., Mertin, J., Brendel, W. Lancet, 1974, ii, 1331. 5. Mertin, J. ibid. p. 717.

1965, 2, 235.

1383 local granuloma as did graft-survival time was 11-0±0’7 days as against 11-2:0-75 days for the animals not treated with naudicelle. These experiments do not support the hypothesis that polyunsaturated fatty acids are specifi-

every

day. This did not produce

L.A. and

oleic acid.

The

a

mean

cally involved in immunregulatory mechanisms. Division of Immunology at the Institute of Physiological Chemistry of the University of Rostock,

Rostock, German Democratic Republic. Institute of Pathology,

Westgate Road, Newcastle upon Tyne NE4 6BE.

J. BROCK. E. J. FIELD.

SATELLITE-STALK STAINING OF HUMAN CHROMOSOMES SiR,—Eiberg 1 claims to have obtained selective Giemsa staining of the satellites of human acrocentric chromosomes following treatment by 0-OlAf NaNOjj or Earle’s BSS medium (pH 7-3) at 87°C for 15 minutes. If this were so the technique would be much simpler than the satellitestaining method of Matsui and Sasaki, which involved Giemsa staining after serial extraction of D.N.A., R.N.A., and acid-soluble proteins. However, we cannot agree with Mr Eiberg The deeply stained spots on the short arms of the acrocentrics after NaN02 treatment are not located on the satellites but on the satellite stalks. This conclusion is based on the

following findings : 1. Dual staining of human chromosomes, first with conventional Giemsa and then with NaNO,-Giemsa, revealed the intensely stained spots on the satellite stalks of the acrocentrics (see figure, a). Pale-stained satellites were occasionally seen on top of the spots. The frequency and size of the spots varied in different acrocentric members, as well as in different individuals -apparently reflecting polymorphism, which may serve as an excellent genetic marker. 2. Dual staining with conventional Giemsa and NaNO.-’ acridine-orange (A.o.) showed bright yellow fluorescent spots on the same sites as those with NaNO,-Giemsa, whereas the satellites, centromere, and arms of the chromosomes appeared orangeThe telomere regions of most chromosomes red (fig., b). fluoresced in the same colour as, but weaker than, the satellite stalks. We have not yet been able to reproduce the telomere bands on black-and-white prints. The banding patterns of NaNO,-A.o. staining were similar to, but more clear-cut than, those of the T bands. 3. We have been using dual-fluorescence staining for Q and T bandsin the study of the origin of chromosome anomalies. A.o. fluorescence after T-band treatment shows marker spots on the satellite stalks of the acrocentrics (fig., c). These T markers are inherited and are found in 10-25% of the acrocentrics, independently of the Q markers which are located on the satellites or paracentromeric regions. In view of the similarities between the T bands and NaNO, bands, it is most likely that both are variants of the same banding patterns. 4. Treatment with Earle’s BSS medium (pH 7-3) at 87°C for 15 minutes, suggested by Mr Eiberg as an alternative to NaNO, treatment, has been known to produce R-banding patterns.’ R banding, which is related to T banding, in our experience also reveals marker spots on the satellite stalks.

These findings indicate that Mr Eiberg’s method stains the satellite stalks of the acrocentric chromosomes. NaNO. banding, however, has some interesting features. The satellite-stalk spots after this treatment are more intense than those with T banding, being clearly visible even on Giemsa staining. The A.o.-stained telomere bands, despite the difficulty of reproducing them photographically, are more pronounced and clear-cut. This feature may be 1. 2. 3. 4.

Eiberg, H. Lancet, 1974, ii, 836. Matsui, S., Sasaki, M. Nature, 1973, 246, 148. Dutrillaux, B. Chromosoma, 1973, 41, 395. Dutrillaux, B., Covic, M. Exp. Cell Res. 1974, 85, 143.

a,

Dual staining of D-group and G-group chromosomes. By conventional Giemsa (left) and Giemsa after NaNO,

treatment (right); NaNO, treatment

b, Giesma (left) and acridine-orange after

c, Q-band (left) and T-band staining (right). Bars indicate satellite stalks (upper) and centromere (lower).

(right);

useful for studying structural anomalies involving the telomere regions. There have been reports of production of T-like banding patterns with A.o. staining after heat treatment with various alkaline solutions.II,8 The pH of 0 OlAf NaNO. is about 9. Therefore, part of its effect is likely due to the

high pH. This work was supported in part by the Ford Foundation Population Programme (64-0411B) and the World Health Organisation. Laboratory of Embryology and Cytogenetics, University Clinic of Gynaecology and Obstetrics, Geneva, Switzerland.

NORIO NIIKAWA TADASHI KAJII.

SIDE-EFFECTS OF PERPHENAZINE SiR,—The toxic side-effects of perphenazine (’Fentazin’) are well established, but I had never witnessed them until recently when a 13-year-old girl, in hospital for manipulation of a knee, was given a single injection of 5 mg. because of an episode of nausea. She had a very severe reaction which she has described in the following essay: " From the beginning I had an idea something was wrong. For after the fentazin injection my leg appeared to be blotchy, red, and slightly swollen. I felt awkwardly dopy and lifeless. Nothing seemed to make sense to me for the rest of that day and I slept for most of the afternoon. Later on that night I had a stiff neck and I was told it was probably the way I had been sleeping. After having my neck rubbed and I had taken two tablets I fell asleep and stayed that way until morning. When I awoke next morning I had another similar act of stiffness, but was given an injection and it slowly went away. " The next spasm of stiffness crept on me very quickly. I became very pale and found my eyes slowly rising to the ceiling and- all I observed were people’s eyebrows and the top of their

5. Chapelle, de la A., Schröder, J., Selander, R., Stenstrand, K. Chromosoma, 1973, 42, 365. 6. Wyandt, H. E., Vlientick, R. F., Magenis, R. E., Hecht, F. Humangenetik, 1974, 23, 119.