- Email: [email protected]
Mo91—AN ALLELE AT THE FIRST HL-A LOCUS
1118
newly isolated type-C R.N.A.
tumour
viruses from
monkeys
and apes. Laboratory of Tumor Cell Biology, National Cancer Institute
have reaffirmed our belief that periodic on sterilisation procedures in hospitals Such checks also make it possible for are still necessary. the laboratory to offer advice on the correct construction of a test pack and on the laundering and drying of towels used in it. The Bowie and Dick test is very valuable in monitoring high-vacuum autoclaves and is probably the most commonly used test of autoclave function, since it has the great advantage that it can be performed quickly and simply by the operator. However, the sensitivity of the test depends completely on that of the tape used to construct the test cross, and accordingly we believe it imperative that each reel of 3 MM 1222 tape should have a batch number on the core of the roll and not merely on the box as at present; if this was done, then reels of a suspect batch could readily be identified. Finally, we should like to emphasise that the Bowie and Dick test is only ancillary to more precise physical measurements, such as thermocouple recordings of temperature and leak-rate tests. These
events
independent checks
Bethesda, Maryland 20014, U.S.A.
ROBERT C. GALLO.
Institute of Cancer Research, College of Physicians and Surgeons of Columbia University, 99 Fort Washington Avenue, New York, New York 10032, U.S.A.
SOL SPIEGELMAN.
ANTRAL-GASTRIN-CELL HYPERPLASIA IN PEPTIC-ULCER DISEASE
SIR,-Firstly, may I draw attention to printing inaccuracies in my letter of May 11 (p. 939)? The reference notation 3 at the end of the fourth paragraph should in fact be 2. The reference 5 on p. 940, together with 4, should The notations 5 and 6 not appear at all at that point. should appear in the third paragraph (2nd sentence, p. 940), and 7 should appear at the end of the fourth paragraph on
Public Health Laboratory, Mytton Oak Road,
p. 940. Finally, could I also take this opportunity to indicate that I did not intend to convey the impression that the a recurrent ulcer should always be vagotomy completion of vagotomy ? What I wish to come over is that we need to recognise in patients with recurrent ulcer those who best require an antrectomy and those for whom
surgery for
Mo91—AN ALLELE AT THE FIRST HL-A LOCUS
or
either vagotomy or vagotomy and antrectomy would be the appropriate procedures. Western General Hospital, Edinburgh EH4 2XU.
W. SIRCUS.
**We apologise to Dr Sircus for introducing this fusion into his references.-ED. L.
con-
C. A. MORRIS P. H. EVERALL.
Shrewsbury, Shropshire.
SiR,—Seignalet 1 reported the finding of an antigen, Mo9l, which behaved as an allele of the first HL-A locus and was characterised by the coexistent positivity of antiHL-A10 and anti-HL-All antibodies. Although Seignalet found an antigen frequency of at least 1 % in a Caucasian population, there seem to have been no further reports of this antigen, and we would like to report another occurrence of Mo9l, discovered during the typing of recipients
awaiting a kidney transplant. BOWIE AND DICK TEST
SIR,-We wish to draw attention to discrepant results obtained with the Bowie and Dick test, which we believe are due to variations in the autoclave tape used. This laboratory has for many years carried out regular checks on autoclaves in the Shrewsbury group of hospitals. As a result of these tests, faults, previously unsuspected by the users, have often been found. Lately we tested a high-vacuum autoclave and showed it to be unserviceable, although the Bowie and Dick test performed by the centralsterile-supply-department (C.S.S.D.) staff a few hours earlier had been satisfactory. The hospital engineer subsequently demonstrated that the autoclave vacuum pump was at fault and was only capable of drawing a vacuum of 27 in. Hg. When this defect was remedied, laboratory and C.S.S.D. tests gave satisfactory results. We had previously offered advice to the staff of the C.S.S.D. on the laundering of towels, preparation of test packs, and performance of the tests, and in fact none of these or similar factors caused the discrepancies. We repeated the tests, using the same autoclave after intentionally inflicting an air leak and also, on another occasion, after the autoclave had developed a natural fault (once again a falling off of pumping efficiency). We found that the sample of 3 MM number 1222 tape used by the laboratory was more sensitive to malfunction of the autoclave (that is, more heat-resistant) than was the batch of tape in daily use in the C.S.S.D. The batch number of the laboratory tape is unknown; that of the C.S.S.D. tape is 2012-2202-0 run no. 6 PIE 3. Perhaps even more disturbing was the fact that even with the more sensitive laboratory tape, a small but nevertheless obvious air leak into an autoclave during the initial vacuum stage was not detected, and a gross leak had to be introduced before the Bowie and Dick test failed.
Our results agree with those of Seignalet in that Mo9l cells react strongly with most HL-A10 antisera (apparently without HL-All antibodies) but react weakly or not at all with many HL-All antisera (apparently without HL-A10 antibodies). The coexistent positivity with HL-A10 and HL-A11antisera is inherited as a single unit in a mendelian fashion, as shown by the following family HL-A genotypes: Mother 1 8
Father
Mo91 W05
A (patient) Mo91 2 7 W05
2 7
2 12
B (brother) Mo91 2 7 W05
C
(brother) 12 8 12
The serological results with the HL-A10 and HL-All antisera were as follows: Serum no. Mother A
25* 60 100 100
B
29
7* Serum no. Mother..
N.T.
A B
N.T.
..
..
*
=
100
45* 100 100 100
86 100 100 100
known oligospecific
HL-AlO (% 108 83 N.T. 100 100 100 N.T. 100
kill) 843*
1221 100 100 80
..
100 100
1252 100
HL-All (% kill) 250 327 566 935 999* 1347 1363 50 100 60 40 20 100 20 100 50 50 100 50 100 50 30 100 20 60 ..
..
sera...
..
=
no
kill.
N.T.
=
not
100 100
1377 100 100 100
tested.
Cells were typed using a modification of the microlymphocytotoxicity technique of Terasaki.2 Lymphocytes were separated on a Ficoll-Triosil gradient.3,4 1 fl.l. cell suspension (1’0-1’5 x 106 cells per ml.) was incubated for 30 minutes with 1 1. antiserum. 5 1. rabbit/human 1. 2. 3. 4.
Seignalet, J. Tissue Antigens, 1972, 2, 454. Terasaki, P. I., McClelland, J. D. Nature, 1964, 204, 998 Boyum, A. Scand. J. clin. Lab. Invest. 1968, 21, suppl. 97. Harris, R., Ukaejiofo, E. Br. J. Hœmat. 1970, 18, 229.
1119 was added and the suspension incubated another 30 minutes. All incubations were at 37°C. Dead cells were stained with trypan-blue. We have not done
complement
absorption or elution studies. All antisera
were
obtained from the National Tissue
Typing
Reference Laboratorv, Bristol. Tissue
Typing Laboratory,
Cardiff Royal Infirmary,
Newport Road, Cardiff CF2 1SZ.
R. B. RUSSELL P. D. BROWN.
ARACHIDONIC ACID AND RETINAL PIGMENTARY DEGENERATION
SIR,-Abnormal lipid metabolism has been associated pigmentary degeneration of the retina in abetalipoproteinacniia (Bassen-Kornzweig disease)and phytanicacid-storage disease (Refsum’s disease).2 Both can be diagnosed by analysis of plasma-lipids. Analysis of plasmafatty-acid levels of a 29-year-old Black woman with pigmentary degeneration of the retina and progressive spinocerebellar degeneration showed the arachidonic-acid (C20:4n-6) level to be 18% (range 15-21%, normal female range 5-9%). Both neutral lipid and phospholipid fractions with
a considerable enrichment in arachidonic-acid content, but triglycerides and esterified cholesterol were primarily affected. The levels could not be modified by altering the patient’s diet.
showed
Fibroblasts cultured from this patient were incubated with [14C]-arachidonate for 24 hours in serum-free medium. These cells incorporated much greater amounts of label into triglyceride, esterified cholesterol, and various phospholipid fractions (especially phosphatidylethanolamine) than PLASMA-ARACHIDONIC-ACID LEVELS IN CONTROLS WITH EYE DISORDERS
AND
PATIENT!
within the reported levels,4 but occasional high levels (up to 11 %) were seen in normal men; this may be related to variations in the level of prostaglandin synthesis, since P.G.E.2 is derived from arachidonic acid.s women with retinitis pigmentosa (a very showed plasma-arachidonic-acid disorder) heterogeneous levels of greater than 9°,0, as did seven out of fifteen female members of their immediate families; in two patients, the level was twice that in normal patients (15% of total plasmafatty-acids). Women with a wide variety of eye disorders (optic atrophy, choroiderma, retinitis pigmentosa associated with diabetes, night blindness) did not show any abnormality in plasma-arachidonic-acid levels, but five women
Six
out
of nine
with uveitis showed a substantial increase and could not be distinguished from the patients with high plasma-arachidonic-acid levels and retinitis pigmentosa. Three out of fourteen male retinitis-pigmentosa patients had plasmaarachidonic-acid levels greater than 13% and six others had levels greater than 9%. Because of the greater variability of plasma-arachidonic-acid in males, studies are in progress to see if analysis of fatty acids in individual lipids will provide clearer differences. We believe that fatty-acid analysis may prove of interest in distinguishing some forms of retinal pigmentary degeneration. The mechanisms controlling the extent of unsaturation of membrane and storage lipid are not well understood, and composition depends upon largely unknown factors controlling competitive enzyme systems governing chain length and degree of unsaturation as well as (to a limited extent) upon diet. Polyunsaturated fatty acids are major constituents of the phospholipids of the outer segments of the retina, and an abnormality in composition may result in degenerative membrane changes. These preliminary studies indicate that a defect in arachidonate metabolism may be involved in certain types of retinitis pigmentosa.
Departments of Pediatrics and
Biochemistry, Department of Ophthalmology, University of Chicago, Chicago, Illinois 60637, U.S.A.
GLYN DAWSON. FRANK W. NEWELL.
HYPONATRÆMIA
did normal human fibroblasts. Incorporation of [14C]linoleate into triglycerides, esterified cholesterol, and phospholipids was also increased compared to controls, whereas [14C]-palmitate or [14C]-stearate incorporation was indistinguishable from normal.
’
, ’
!I
Arachidonic acid is derived by desaturation and chain elongation from linoleic acid (Cis:2n-6), an essential fatty acid derived mainly from plant sources; unsaturated fatty acids (especially C22:6n-6) are present in high concentration in retinal outer segment lipids.33 This apparent abnormality of arachidonic-acid metabolism prompted a survey of plasma-arachidonate levels in patients with pigmentary degeneration of the retina and their immediate families, patients with a variety of retinal diseases, and fifty patients with unrelated disorders (table). In women, the normal arachidonic-acid level was 1. 2. 3.
Bassen, F. A., Kornzweig, A. L. Blood, 1950, 5, 381. Refsum, S. Acta psychiat. scand. 1946, 38, 9. Borggreven, J. M. P. M., Daeman, F. J. M., Bonting, S. L. Biochim. biophys. Acta, 1970, 202, 374.
SIR,-Your editorial (March 2, p. 342) seems to perpetuate the misconception that " sick cells " somehow are responsible for hyponatraemia. The view presented that sodium from the extracellular fluid enters suggests " sick " cells and that the cell membranes of such cells are so leaky that normally non-diffusable intracellular solutes leave the cell. To the extent that such solute represents an equiosmolar exchange for extracellular fluid, the serumsodium concentration could fall without a corresponding drop in the serum osmolality. Such, however, is not the case in clinical states of hyponatraemia in which both serum-sodium and osmolality are proportionately reduced 6-8. Most intracellular non-diffusable molecules are of fairly high molecular weight; however, if molecules as small as 200 daltons left these " sick cells " in exchange for sodium their combined concentrations in plasma would have to increase to 800 mg. per 100 ml. to compensate for a drop in serum-sodium from 140 to 120 meq. per 1. Perhaps in rapidly changing agonal states with loss of renal function such a situation may occur transiently, but this is not the 4. Kirkeby, K., Bjerkedal, I. Acta med. scand. 1968, 183, 143. 5. Samuelsson, B. E. Fedn Proc. 1972, 31, 1442. 6. Leaf, A., Mamby, A. R. J. clin. Invest. 1952, 31, 60. 7. Edelman, I. S., Leibman, J., O’Meara, M. P., Birkenfeld, L. W. ibid. 1958, 37, 1236. 8. Leaf, A. New Engl. J. Med. 1962, 267, 3.
newly isolated type-C R.N.A.
tumour
viruses from
monkeys
and apes. Laboratory of Tumor Cell Biology, National Cancer Institute
have reaffirmed our belief that periodic on sterilisation procedures in hospitals Such checks also make it possible for are still necessary. the laboratory to offer advice on the correct construction of a test pack and on the laundering and drying of towels used in it. The Bowie and Dick test is very valuable in monitoring high-vacuum autoclaves and is probably the most commonly used test of autoclave function, since it has the great advantage that it can be performed quickly and simply by the operator. However, the sensitivity of the test depends completely on that of the tape used to construct the test cross, and accordingly we believe it imperative that each reel of 3 MM 1222 tape should have a batch number on the core of the roll and not merely on the box as at present; if this was done, then reels of a suspect batch could readily be identified. Finally, we should like to emphasise that the Bowie and Dick test is only ancillary to more precise physical measurements, such as thermocouple recordings of temperature and leak-rate tests. These
events
independent checks
Bethesda, Maryland 20014, U.S.A.
ROBERT C. GALLO.
Institute of Cancer Research, College of Physicians and Surgeons of Columbia University, 99 Fort Washington Avenue, New York, New York 10032, U.S.A.
SOL SPIEGELMAN.
ANTRAL-GASTRIN-CELL HYPERPLASIA IN PEPTIC-ULCER DISEASE
SIR,-Firstly, may I draw attention to printing inaccuracies in my letter of May 11 (p. 939)? The reference notation 3 at the end of the fourth paragraph should in fact be 2. The reference 5 on p. 940, together with 4, should The notations 5 and 6 not appear at all at that point. should appear in the third paragraph (2nd sentence, p. 940), and 7 should appear at the end of the fourth paragraph on
Public Health Laboratory, Mytton Oak Road,
p. 940. Finally, could I also take this opportunity to indicate that I did not intend to convey the impression that the a recurrent ulcer should always be vagotomy completion of vagotomy ? What I wish to come over is that we need to recognise in patients with recurrent ulcer those who best require an antrectomy and those for whom
surgery for
Mo91—AN ALLELE AT THE FIRST HL-A LOCUS
or
either vagotomy or vagotomy and antrectomy would be the appropriate procedures. Western General Hospital, Edinburgh EH4 2XU.
W. SIRCUS.
**We apologise to Dr Sircus for introducing this fusion into his references.-ED. L.
con-
C. A. MORRIS P. H. EVERALL.
Shrewsbury, Shropshire.
SiR,—Seignalet 1 reported the finding of an antigen, Mo9l, which behaved as an allele of the first HL-A locus and was characterised by the coexistent positivity of antiHL-A10 and anti-HL-All antibodies. Although Seignalet found an antigen frequency of at least 1 % in a Caucasian population, there seem to have been no further reports of this antigen, and we would like to report another occurrence of Mo9l, discovered during the typing of recipients
awaiting a kidney transplant. BOWIE AND DICK TEST
SIR,-We wish to draw attention to discrepant results obtained with the Bowie and Dick test, which we believe are due to variations in the autoclave tape used. This laboratory has for many years carried out regular checks on autoclaves in the Shrewsbury group of hospitals. As a result of these tests, faults, previously unsuspected by the users, have often been found. Lately we tested a high-vacuum autoclave and showed it to be unserviceable, although the Bowie and Dick test performed by the centralsterile-supply-department (C.S.S.D.) staff a few hours earlier had been satisfactory. The hospital engineer subsequently demonstrated that the autoclave vacuum pump was at fault and was only capable of drawing a vacuum of 27 in. Hg. When this defect was remedied, laboratory and C.S.S.D. tests gave satisfactory results. We had previously offered advice to the staff of the C.S.S.D. on the laundering of towels, preparation of test packs, and performance of the tests, and in fact none of these or similar factors caused the discrepancies. We repeated the tests, using the same autoclave after intentionally inflicting an air leak and also, on another occasion, after the autoclave had developed a natural fault (once again a falling off of pumping efficiency). We found that the sample of 3 MM number 1222 tape used by the laboratory was more sensitive to malfunction of the autoclave (that is, more heat-resistant) than was the batch of tape in daily use in the C.S.S.D. The batch number of the laboratory tape is unknown; that of the C.S.S.D. tape is 2012-2202-0 run no. 6 PIE 3. Perhaps even more disturbing was the fact that even with the more sensitive laboratory tape, a small but nevertheless obvious air leak into an autoclave during the initial vacuum stage was not detected, and a gross leak had to be introduced before the Bowie and Dick test failed.
Our results agree with those of Seignalet in that Mo9l cells react strongly with most HL-A10 antisera (apparently without HL-All antibodies) but react weakly or not at all with many HL-All antisera (apparently without HL-A10 antibodies). The coexistent positivity with HL-A10 and HL-A11antisera is inherited as a single unit in a mendelian fashion, as shown by the following family HL-A genotypes: Mother 1 8
Father
Mo91 W05
A (patient) Mo91 2 7 W05
2 7
2 12
B (brother) Mo91 2 7 W05
C
(brother) 12 8 12
The serological results with the HL-A10 and HL-All antisera were as follows: Serum no. Mother A
25* 60 100 100
B
29
7* Serum no. Mother..
N.T.
A B
N.T.
..
..
*
=
100
45* 100 100 100
86 100 100 100
known oligospecific
HL-AlO (% 108 83 N.T. 100 100 100 N.T. 100
kill) 843*
1221 100 100 80
..
100 100
1252 100
HL-All (% kill) 250 327 566 935 999* 1347 1363 50 100 60 40 20 100 20 100 50 50 100 50 100 50 30 100 20 60 ..
..
sera...
..
=
no
kill.
N.T.
=
not
100 100
1377 100 100 100
tested.
Cells were typed using a modification of the microlymphocytotoxicity technique of Terasaki.2 Lymphocytes were separated on a Ficoll-Triosil gradient.3,4 1 fl.l. cell suspension (1’0-1’5 x 106 cells per ml.) was incubated for 30 minutes with 1 1. antiserum. 5 1. rabbit/human 1. 2. 3. 4.
Seignalet, J. Tissue Antigens, 1972, 2, 454. Terasaki, P. I., McClelland, J. D. Nature, 1964, 204, 998 Boyum, A. Scand. J. clin. Lab. Invest. 1968, 21, suppl. 97. Harris, R., Ukaejiofo, E. Br. J. Hœmat. 1970, 18, 229.
1119 was added and the suspension incubated another 30 minutes. All incubations were at 37°C. Dead cells were stained with trypan-blue. We have not done
complement
absorption or elution studies. All antisera
were
obtained from the National Tissue
Typing
Reference Laboratorv, Bristol. Tissue
Typing Laboratory,
Cardiff Royal Infirmary,
Newport Road, Cardiff CF2 1SZ.
R. B. RUSSELL P. D. BROWN.
ARACHIDONIC ACID AND RETINAL PIGMENTARY DEGENERATION
SIR,-Abnormal lipid metabolism has been associated pigmentary degeneration of the retina in abetalipoproteinacniia (Bassen-Kornzweig disease)and phytanicacid-storage disease (Refsum’s disease).2 Both can be diagnosed by analysis of plasma-lipids. Analysis of plasmafatty-acid levels of a 29-year-old Black woman with pigmentary degeneration of the retina and progressive spinocerebellar degeneration showed the arachidonic-acid (C20:4n-6) level to be 18% (range 15-21%, normal female range 5-9%). Both neutral lipid and phospholipid fractions with
a considerable enrichment in arachidonic-acid content, but triglycerides and esterified cholesterol were primarily affected. The levels could not be modified by altering the patient’s diet.
showed
Fibroblasts cultured from this patient were incubated with [14C]-arachidonate for 24 hours in serum-free medium. These cells incorporated much greater amounts of label into triglyceride, esterified cholesterol, and various phospholipid fractions (especially phosphatidylethanolamine) than PLASMA-ARACHIDONIC-ACID LEVELS IN CONTROLS WITH EYE DISORDERS
AND
PATIENT!
within the reported levels,4 but occasional high levels (up to 11 %) were seen in normal men; this may be related to variations in the level of prostaglandin synthesis, since P.G.E.2 is derived from arachidonic acid.s women with retinitis pigmentosa (a very showed plasma-arachidonic-acid disorder) heterogeneous levels of greater than 9°,0, as did seven out of fifteen female members of their immediate families; in two patients, the level was twice that in normal patients (15% of total plasmafatty-acids). Women with a wide variety of eye disorders (optic atrophy, choroiderma, retinitis pigmentosa associated with diabetes, night blindness) did not show any abnormality in plasma-arachidonic-acid levels, but five women
Six
out
of nine
with uveitis showed a substantial increase and could not be distinguished from the patients with high plasma-arachidonic-acid levels and retinitis pigmentosa. Three out of fourteen male retinitis-pigmentosa patients had plasmaarachidonic-acid levels greater than 13% and six others had levels greater than 9%. Because of the greater variability of plasma-arachidonic-acid in males, studies are in progress to see if analysis of fatty acids in individual lipids will provide clearer differences. We believe that fatty-acid analysis may prove of interest in distinguishing some forms of retinal pigmentary degeneration. The mechanisms controlling the extent of unsaturation of membrane and storage lipid are not well understood, and composition depends upon largely unknown factors controlling competitive enzyme systems governing chain length and degree of unsaturation as well as (to a limited extent) upon diet. Polyunsaturated fatty acids are major constituents of the phospholipids of the outer segments of the retina, and an abnormality in composition may result in degenerative membrane changes. These preliminary studies indicate that a defect in arachidonate metabolism may be involved in certain types of retinitis pigmentosa.
Departments of Pediatrics and
Biochemistry, Department of Ophthalmology, University of Chicago, Chicago, Illinois 60637, U.S.A.
GLYN DAWSON. FRANK W. NEWELL.
HYPONATRÆMIA
did normal human fibroblasts. Incorporation of [14C]linoleate into triglycerides, esterified cholesterol, and phospholipids was also increased compared to controls, whereas [14C]-palmitate or [14C]-stearate incorporation was indistinguishable from normal.
’
, ’
!I
Arachidonic acid is derived by desaturation and chain elongation from linoleic acid (Cis:2n-6), an essential fatty acid derived mainly from plant sources; unsaturated fatty acids (especially C22:6n-6) are present in high concentration in retinal outer segment lipids.33 This apparent abnormality of arachidonic-acid metabolism prompted a survey of plasma-arachidonate levels in patients with pigmentary degeneration of the retina and their immediate families, patients with a variety of retinal diseases, and fifty patients with unrelated disorders (table). In women, the normal arachidonic-acid level was 1. 2. 3.
Bassen, F. A., Kornzweig, A. L. Blood, 1950, 5, 381. Refsum, S. Acta psychiat. scand. 1946, 38, 9. Borggreven, J. M. P. M., Daeman, F. J. M., Bonting, S. L. Biochim. biophys. Acta, 1970, 202, 374.
SIR,-Your editorial (March 2, p. 342) seems to perpetuate the misconception that " sick cells " somehow are responsible for hyponatraemia. The view presented that sodium from the extracellular fluid enters suggests " sick " cells and that the cell membranes of such cells are so leaky that normally non-diffusable intracellular solutes leave the cell. To the extent that such solute represents an equiosmolar exchange for extracellular fluid, the serumsodium concentration could fall without a corresponding drop in the serum osmolality. Such, however, is not the case in clinical states of hyponatraemia in which both serum-sodium and osmolality are proportionately reduced 6-8. Most intracellular non-diffusable molecules are of fairly high molecular weight; however, if molecules as small as 200 daltons left these " sick cells " in exchange for sodium their combined concentrations in plasma would have to increase to 800 mg. per 100 ml. to compensate for a drop in serum-sodium from 140 to 120 meq. per 1. Perhaps in rapidly changing agonal states with loss of renal function such a situation may occur transiently, but this is not the 4. Kirkeby, K., Bjerkedal, I. Acta med. scand. 1968, 183, 143. 5. Samuelsson, B. E. Fedn Proc. 1972, 31, 1442. 6. Leaf, A., Mamby, A. R. J. clin. Invest. 1952, 31, 60. 7. Edelman, I. S., Leibman, J., O’Meara, M. P., Birkenfeld, L. W. ibid. 1958, 37, 1236. 8. Leaf, A. New Engl. J. Med. 1962, 267, 3.