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renal or cardiac complications (mean values of pretreatmentsystolic and diastolic blood-pressure, 213/121 mm Hg), the fallin blood-pressure induced by intravenous injection of clonidine(0.15 mg) was accompanied by a significant increase in plas-ma-prolactin. The maximum level was reached at the timewhen the blood-pressure was lowest, 30 min after the drug wasgiven (see figure). In our patients the basal prolactin concent-ration was normal while in the patients described by ProfessorStumpe and colleagues (July 30, p. 211) it was abnormallyhigh. This difference may have arisen because we studied only4 patients; some of the patients studied by Stumpe et- al. hadhad a normal plasma-prolactin. These authors also found thatoral bromocriptine had an antihypertensive effect which wasfollowed by a decrease of prolactin to normal. Bromocriptineinduced a slow fall in blood-pressure while clonidine induceda sudden fall. Clonidine seems to exert stress evoking a reflexrise in prolactin, but how this occurs-for example, whetherthere is a decrease in hypothalamic dopaminergic tone-is notknown.

Institutes of Pharmacology andMedical Pathology,

University of Messina,Messina, Italy

G. NISTICO’F. TRIMARCHIF. CONSOLO

UTILISATION OF DISCARDED MEDIUM INAMNIOTIC-FLUID CELL CULTURES

SiR,—The value of prenatal diagnosis of genetic disordersis now well recognised, and one of the essential elements of aprenatal diagnostic programme is laboratory performance.Where amniotic-fluid cells are needed to make a diagnosis, suc-cessful culture is important. Success depends on the number ofviable cells that adhere to the culture flasks. Too little fluidand/or too few cells lessens the chance of successful culture.This disadvantage can be overcome by replanting the superna-tant medium which is usually discarded. The procedure yieldsan adequate number of cells for analysis and obviates the needfor a repeat amniocentesis.

Amniotic fluid obtained from the second-trimester preg-nancy was centrifuged (200 g) for 10 main and the cell buttonwas resuspended in 1-2 ml of fetal calf serum. The cells werethen planted in 25 cm2 Falcon flasks and placed in a 5% CO2incubator at 37 °C for 15-30 min. McCoy’s SA medium (5 ml)enriched with 20% fetal calf serum were added, and the flaskswere left undisturbed in the incubator. After 4-5 days; super-natant medium of the culture was poured off and, instead ofbeing discarded, was replanted into another flask to which anequal amount of fresh medium has been added. Fresh mediumwas also added to the original flask where a few cells had set-tled to the bottom. Thus, an additional flask of culture wasmade available.

’

- -

"Pour-ofT’ medium can also -be used when a culture isready for chromosome harvest in situ. The procedure is essen-tially the same as described above. We believe that the cellsthat are reseeded are the loosely attached mitotic cells’ whichare being transferred in the supernatant medium. - -

In our experience pour-off medium in amniotic-fluid cell cul-ture can be used to establish viable cultures for at least fivesuccessive times. G-banded karyotypes prepared for the fifthpour-off culture were identical to the original karyotypes.We now routinely use this pour-off method in prenatal chro-

mosome diagnosis to ensure that enough cells are available foranalysis. The procedure is especially useful in complex cyto-genetic conditions, because it provides more cells for bandingstudies.

Cellular Genetics Laboratory,Pathology Division,Bureau of Laboratories,Center for Disease Control,Atlanta, Georgia 30333, U.S.A.

SUZANNE TRUSLER

JANICE BULLARDANDREW T. L. CHEN

7. Noel, G. L., Suh, H. K., Gilbert-Stone, J., Frants, A. L. G. J. clin. Endocr.Metab. 1972, 35, 840.

1. Terasima, T., Tolmach, L. J. Expl. Cell Res. 1963, 30, 344.

ALCOHOL AND HYPERTENSION

SIR,-Recent reports’ have suggested an associationbetween alcohol intake and raised blood-pressure, and in 1966Klatsky et al. suggested that three or more alcoholic drinkstaken every day is a risk factor for hypertension. These reportsdo not refer to nutritional status and its possible associationwith hypertension among alcoholics.We have studied 61 alcoholic patients with pellagra aged

23-49, most of them drinking more than 800 ml of pinga (aBrazilian alcoholic beverage similar to white rum) containingabout 300 ml of pure ethanol.

Besides the typical skin lesions of pellagra, 31 patients haddiarrhcea and 27 had peripheral neuropathy. Low haemoglobinand serum-albumin values were found in 25 and 36 patients,respectively. The diastolic blood-pressure was above 95 mmHg in 44 pauents. ,

The diastolic blood-pressure came down while the patientswere in hospital, even without the administration of hypoten-sive drugs;4 30 of the 44 hypertensive patients had normaldiastolic blood-pressures when they were discharged.

Leonard et al. suggest an increase in corticosteroids as a

possible factor for hypertension in alcoholism. It is also pos-sible that the renin-angiotensin system is stimulated bydeficiency of certain B vitamins, notably pyridoxine.6 Inkwashiorkor urinary excretion of catecholamine metabolites isincreased, and this could also be a factor in the aaiology ofhypertension associated with the nutritional status of the alco-holics.’ Colleagues in.the department of pathology8 have foundhigh levels of heart-muscle catecholamines in chronic alcoholicrats.

We are now looking for a relation between tryptophan/nia-cin/serotonin metabolism and hypertension.

Department of Medicine,Faculty of Medicine,Botucatu and Ribeirão Preto,São Paulo, Brazil

HÉLIO VANNUCCHIALVARO OSCAR CAMPANAJOSÉ EDUARDO DUTRA DE OLIVEIRA

FAILURE OF ANTI-THYMOCYTE GLOBULINTREATMENT IN T-CELL LYMPHOMA

SIR,—Dr Edelson and his colleagues (July 30, p. 249) suc-cessfully treated mycosis fungoides with an’anti-thymocyte glo-bulin. We have had very different results. Our patient, a75-year-old female, had a 15-year history of mycosis fungoideswhich had been contained by local electron beam therapy andtopical nitrogen mustard. The disease escaped control andreached the tumour stage, confirmed by biopsy, with regionallymph-node involvement. The white-blood-cell count was low(3200/µl total; 650/µl lymphocytes; 580/µl T lymphocytes,measured by rosetting with sheep erythrocytes). The patientwas treated with anti-thymocyte globulin ’Pressimmune’

(Hoechst Pharmaceuticals) exactly as described by Edelson etal. After 10 days the T cells were reduced to 250/µl and thetotal lymphocytes to 550/µl. There was no serum sickness andno methylprednisolone was required. However, the patient’scondition deteriorated and the existing tumours continued toenlarge. After 3 weeks, when several new ulcerating tumourshad developed, treatment was abandoned.The use of anti-thymocyte serum is based on evidence that

the tumour stage of mycosis fungoides is a T-cell lymphoma.In our patient, treatment was ineffective and possibly deleter-

1. Ramsey, L. E. Lancet, 1977, ii, 111.2. Beevers, D. G. ibid. p. 114.3. Klatsky, A. L., Friedman, G. D., Siezelaub, A. B., Gerard, M. J. New Engl.

J. Med. 1966, 296, 1194.4. Vannucchi, H., Campana, A. O. Arq. Gastroent. São Paulo, 1974, 11, 129.5. Leonard, J. W., Gilford, R., Humphrey, D. C. Am. Heart J. 1965, 69, 610.6. De Lorme, C. B., Lupien, P. J., Despointes, R. H. J. Nutr. 1975, 105, 1192.7. Hoeldtke, R. D., Wurtman, R. J. Am. J. clin. Nutr. 1973, 26, 205.8. Rossi, M. A., Oliveira, J. S. M., Zucoloto, S., Becker, P. F. L. Beitr. Path.

Bd. 1976, 159, 51.