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Salivary secretion of ABH blood groupsubstances in cystic fibrosis of the pancreas
Volume 68
Number 1
Brie[ clinical and laboratory observations
received the standard diet plus 0.6 Gm. pet" kilogram per day orotic acid via a gastric tube. T h e fourth group received the high galactose diet (25 per cent) and 0.6 Gin. per kilogram orotic acid daily. T h e lenses were examined by biomicroscopy every third day to observe the possibility of cataract development. Blood studies were performed after 22 days to estimate levels of cholesterol, lipids, plasma proteins, and lipoproteins. Galactose in blood and urine was detected by chromatography. The liver, kidneys, pancreas, eyes, and bones were examined histologically. RESULTS
The rats in the control group had no abnormal biochemical or histologic changes. T h e animals receiving the galactose-rich diet had galactosuria, galactosemia, and cataracts, but there were no changes in the blood values of cholesterol, lipids, and protein, and no histologic changes in the liver, kidneys, pancreas, and bones. T h e animals which received the orotic acid and the standard diet had no abnormal biochemical or histologic changes. T h e biologic and histologic changes in the rats fed the galactose-rich diet together with orotic acid were similar to those observed in the rats of Group I I which did not receive orotic acid.
139
CONCLUSIONS O u r observations do not indicate that orotic acid modifies the noxious effects of a galactose-rich diet in rats nor do they provide evidence of toxicity within the limits of the dose employed. REFERENCES
1. Tada, K., Kydo, Z., Ohno, T., Akabane, J., and Chiba, R.: Congenital galactosemia and orotic acid therapy with promising results: preliminary report, Tohoku J. Exper. Med. 77: 340, 1962. 2. Tada, K., Akabane, J., and Yokoyarna, M.: Beneficial effect of orotic acid upon galactose metabolism of erythrocytes from congenital galactosemia, Tohoku J. Exper. Med. 77: 400, 1962. 3. Standerfer, S. M., and Handler, P.: Fatty liver induced by orotic acid feeding, Proc. Soc. Exper. Biol. & Med. 90: 270, 1955. 4. Handschumacher, R. F., Creasey, W. A., Jaffe, J. J., Pasternak, C. A., and Hankin, L.: Biochemical and nutritional studies on the induction of fatty livers by dietary orotic acid, Proc. Nat. Acad. Sc. 46: 178, 1960. 5. Suva, J., Cerhova, M., Habermannova, S., and Habermann, V.: Inhibitory effect of 6-Azauracil on the induction of fatty liver by dietary orotic acid in young rats, Acta vitamin. 15: 97, 1961. 6. Creasey, W. A., Hankin, L., and I-Iandschureacher, R. F.: Fatty livers induced by orotie acid. I. Accumulation and metabolism of lipids, J. Biol. Chem. 236: 2064, 1961. 7. Sidransky, H., Verney, E., and Lombardi, B.: Factors influencing the induction of fatty liver by orotic acid, J. Nutrition 81: 348, 1963.
Salivary secretion of ABH blood group substances in cystic fibrosis of the pancreas S i m o Virtanen, M . D . "x" B E T I I E S D A , M D . , AND W A S I I 1 N G T O N , D.C.
From the Pediatric Metabolism Branch, National Institutes o[ Health, Bethesda, and Children's Hospital, Washington, D. C. Address, c/o Paul A. di Sant'Agnese, M.D., Chief Pediatric Metabolism Branch, National Institutes o[ Health, Bethesda, Md. ~Fellow of the National Cystic Fibrosis Research Foundation.
C Y s T I c F I B R O S I S is a generalized hereditary disease in which there is a dysfunction of many exocrine glands. U p to the present there has been no convincing evidence for linkage or association between
1 40
January 1966
Brie[ clinical and laboratory observations
T a b l e I. T e c h n i q u e
A2 cells + 4 HA units of anti-A B cells + 4 HA units of anti-B O cells + 4 HA units of anti-H
Dilution 23 2a 23
22 22 22
o[ saliva 24 ~ 219 2'~ ~ 219 2~ ---> 219
I
1 1 1
Controls 2 3 2 3 2 3
4 4 4
Controls 1, Cells + saline. 2, Cells + 4 H A units of antiserum. 3, Cells + 2 H A units o[ antiserum. 4, Cells + sMiva diluted 8 9
cystic fibrosis and any of tile n u m e r o u s genes tested. Several studies have demonstrated the i n v o l v e m e n t of salivary glands in cystic fibrosis? I n particular, the concentration of fucose has been shown to be higher in saliva from patients with cystic fibrosis t h a n in saliva from n o r m a l control subjects? Lfucose is one of the m a j o r sugar components of blood group substances a n d the H-specificity is associated with L-fucose. 2 Accordingly, it seemed reasonable to study the possible association between secretor status a n d cystic fibrosis a n d the titer of blood group substances secreted in saliva of patients with cystic fibrosis. MATERIAL
AND
METHODS
T h e salivary secretion of ABH blood group substances was determined quantitatively in 65 patients with cystic fibrosis a n d in 40 r a n d o m children and adult control subjects. Saliva was collected utilizing a r u b b e r b a n d as the stimulus for salivation. T h e samples of saliva were immediately heated in a boiling water b a t h for 10 minutes, centrifuged, a n d the s u p e r n a t a n t kept at - 2 0 ~ C. u n t i l tested. Each sample was titrated with anti-A a n d anti-B sera a n d a n t i - H (Ulex extract) using A2, B, or O cells, respectively. Anti-A a n d anti-B sera were obtained commercially ( O r t h o ) . A n t i - H reagent was prepared by the author. T h e test was performed in 12 x 75 ram. test tubes at room temperature: as follows: Saliva was diluted with antiserum c o n t a i n i n g 4 hemagglutinatifig units. A n equal a m o u n t of 1 per cent red cell suspension was added a n d the tubes were well shaken. T h e total
T a b l e II. Distribution of secretors a n d nonsecretors Secretors No.
Per cent
49 31
76.9 77.0
CF patients Control subjects
secretors NonPer No. cent
16 9
23.1 23.0
Total No.
65 40
T a b l e I I I . T i t e r of group-specific substances in saliva CF patients Substance
No. t Mean titer
A A+H B B+H A+B+H H
1 18 1 3 3 23 49
volume in developed read after the details
0.82 x 3.34x 0.64 x 4.64x 3.07x 1.7 x
104 104 102 104 10:~ 10~
. Control subjects No. I Mean titer
2 14 3 12
1.64x 1.4x 2.73 x 1.61 x
104 104 10* 102
31
the tubes was 0.5 ml. T h e patterns at the bottom of the tubes were 2 hours a n d 4 hours. T a b l e I lists of the procedure.
RESULTS
T a b l e I I lists the frequencies of secretors a n d nonsecretors. Percentages for patients with cystic fibrosis a n d for control subjects are essentially the same. These frequencies did n o t differ from those reported for the n o r m a l population: 76 to 78 per cent secretors and 22 to 24 per cent nonsecretors. 2, 3 I n addition, no significant difference was f o u n d between titers of the group-specific
Volume 68
Number 1
substances in saliva studied ( T a b l e I I I ) .
Brief clinical and laboratory observations
of
the
two
groups
SUMMARY T h e salivary secretion of A B H blood group substances was studied in 65 patients with cystic fibrosis a n d 40 n o r m a l control subjects. No association was d e m o n s t r a t e d between A B H secretory status and cystic fibrosis.
14 l
REFERENCES 1. Chernick, W. S., and Barbero, G. J.: Studies on human tracheobronchial and submaxillary secretions in normal and pathophysiological conditions, Ann. New York Acad. Sc. 106: 698, 1963. 2. Race, R. R., and Sanger, R.: Blood groups in man, Oxford, 1962, Blackwell Scientific Publications. 3. Wiener, A. S.: Blood groups and transfusion, New York, 1962, Hafner Publishing Company, Inc.
A modified electrode and lead assembly for rectorcardiograpby in newborn and small infants Lorin E. Ainger, M.D., "x"a n d Bill R. C a m p b e l l MEMPHIS,
TENN.
T w o o v the most difficult technical problems of e l e c t r o c a r d i o g r a p h y a n d vectorcard i o g r a p h y in n e w b o r n a n d small infants are m a i n t e n a n c e of firm sldn-to-electrode contact a n d m a i n t e n a n c e of secure lead-electrode connections. A disposable adhesive electrode with snap c o n n e c t o r t p r o v i d e d a satisfacto W solution to these problems. This electrode is a 5//s inch square, silver impregn a t e d meshed cloth fixed to a 11~ inch square plastic adhesive b a n d a g e with an o r d i n a r y g r i p - s n a p p e r to w h i c h the lead is securely fastened. T h e cost of this electrode, however, p r e c l u d e d its use on a large scale; therefore, From the section o/Pediatric Cardiolog~, Department of Pediatrics, University of Tennessee College of Medicine. This work was supported by Grant No. HE-5755-C3, National Heart Institute, National Institutes of Health, United States Public Health Service. "X'Address, 860 Madison Avenue, Memphis, Tenn. 38103. tTelemedies, Inc., Southampton, Pa.
Fig. 1. Components of the modified electrode and lead assembly. From left to right: (1) the electrode, 5/8" in diameter; (2) the 1 ~ " square plastic adhesive bandage with the hole punched in the center; (3) the lead with the connector to fit the grip-snapper.
Number 1
Brie[ clinical and laboratory observations
received the standard diet plus 0.6 Gm. pet" kilogram per day orotic acid via a gastric tube. T h e fourth group received the high galactose diet (25 per cent) and 0.6 Gin. per kilogram orotic acid daily. T h e lenses were examined by biomicroscopy every third day to observe the possibility of cataract development. Blood studies were performed after 22 days to estimate levels of cholesterol, lipids, plasma proteins, and lipoproteins. Galactose in blood and urine was detected by chromatography. The liver, kidneys, pancreas, eyes, and bones were examined histologically. RESULTS
The rats in the control group had no abnormal biochemical or histologic changes. T h e animals receiving the galactose-rich diet had galactosuria, galactosemia, and cataracts, but there were no changes in the blood values of cholesterol, lipids, and protein, and no histologic changes in the liver, kidneys, pancreas, and bones. T h e animals which received the orotic acid and the standard diet had no abnormal biochemical or histologic changes. T h e biologic and histologic changes in the rats fed the galactose-rich diet together with orotic acid were similar to those observed in the rats of Group I I which did not receive orotic acid.
139
CONCLUSIONS O u r observations do not indicate that orotic acid modifies the noxious effects of a galactose-rich diet in rats nor do they provide evidence of toxicity within the limits of the dose employed. REFERENCES
1. Tada, K., Kydo, Z., Ohno, T., Akabane, J., and Chiba, R.: Congenital galactosemia and orotic acid therapy with promising results: preliminary report, Tohoku J. Exper. Med. 77: 340, 1962. 2. Tada, K., Akabane, J., and Yokoyarna, M.: Beneficial effect of orotic acid upon galactose metabolism of erythrocytes from congenital galactosemia, Tohoku J. Exper. Med. 77: 400, 1962. 3. Standerfer, S. M., and Handler, P.: Fatty liver induced by orotic acid feeding, Proc. Soc. Exper. Biol. & Med. 90: 270, 1955. 4. Handschumacher, R. F., Creasey, W. A., Jaffe, J. J., Pasternak, C. A., and Hankin, L.: Biochemical and nutritional studies on the induction of fatty livers by dietary orotic acid, Proc. Nat. Acad. Sc. 46: 178, 1960. 5. Suva, J., Cerhova, M., Habermannova, S., and Habermann, V.: Inhibitory effect of 6-Azauracil on the induction of fatty liver by dietary orotic acid in young rats, Acta vitamin. 15: 97, 1961. 6. Creasey, W. A., Hankin, L., and I-Iandschureacher, R. F.: Fatty livers induced by orotie acid. I. Accumulation and metabolism of lipids, J. Biol. Chem. 236: 2064, 1961. 7. Sidransky, H., Verney, E., and Lombardi, B.: Factors influencing the induction of fatty liver by orotic acid, J. Nutrition 81: 348, 1963.
Salivary secretion of ABH blood group substances in cystic fibrosis of the pancreas S i m o Virtanen, M . D . "x" B E T I I E S D A , M D . , AND W A S I I 1 N G T O N , D.C.
From the Pediatric Metabolism Branch, National Institutes o[ Health, Bethesda, and Children's Hospital, Washington, D. C. Address, c/o Paul A. di Sant'Agnese, M.D., Chief Pediatric Metabolism Branch, National Institutes o[ Health, Bethesda, Md. ~Fellow of the National Cystic Fibrosis Research Foundation.
C Y s T I c F I B R O S I S is a generalized hereditary disease in which there is a dysfunction of many exocrine glands. U p to the present there has been no convincing evidence for linkage or association between
1 40
January 1966
Brie[ clinical and laboratory observations
T a b l e I. T e c h n i q u e
A2 cells + 4 HA units of anti-A B cells + 4 HA units of anti-B O cells + 4 HA units of anti-H
Dilution 23 2a 23
22 22 22
o[ saliva 24 ~ 219 2'~ ~ 219 2~ ---> 219
I
1 1 1
Controls 2 3 2 3 2 3
4 4 4
Controls 1, Cells + saline. 2, Cells + 4 H A units of antiserum. 3, Cells + 2 H A units o[ antiserum. 4, Cells + sMiva diluted 8 9
cystic fibrosis and any of tile n u m e r o u s genes tested. Several studies have demonstrated the i n v o l v e m e n t of salivary glands in cystic fibrosis? I n particular, the concentration of fucose has been shown to be higher in saliva from patients with cystic fibrosis t h a n in saliva from n o r m a l control subjects? Lfucose is one of the m a j o r sugar components of blood group substances a n d the H-specificity is associated with L-fucose. 2 Accordingly, it seemed reasonable to study the possible association between secretor status a n d cystic fibrosis a n d the titer of blood group substances secreted in saliva of patients with cystic fibrosis. MATERIAL
AND
METHODS
T h e salivary secretion of ABH blood group substances was determined quantitatively in 65 patients with cystic fibrosis a n d in 40 r a n d o m children and adult control subjects. Saliva was collected utilizing a r u b b e r b a n d as the stimulus for salivation. T h e samples of saliva were immediately heated in a boiling water b a t h for 10 minutes, centrifuged, a n d the s u p e r n a t a n t kept at - 2 0 ~ C. u n t i l tested. Each sample was titrated with anti-A a n d anti-B sera a n d a n t i - H (Ulex extract) using A2, B, or O cells, respectively. Anti-A a n d anti-B sera were obtained commercially ( O r t h o ) . A n t i - H reagent was prepared by the author. T h e test was performed in 12 x 75 ram. test tubes at room temperature: as follows: Saliva was diluted with antiserum c o n t a i n i n g 4 hemagglutinatifig units. A n equal a m o u n t of 1 per cent red cell suspension was added a n d the tubes were well shaken. T h e total
T a b l e II. Distribution of secretors a n d nonsecretors Secretors No.
Per cent
49 31
76.9 77.0
CF patients Control subjects
secretors NonPer No. cent
16 9
23.1 23.0
Total No.
65 40
T a b l e I I I . T i t e r of group-specific substances in saliva CF patients Substance
No. t Mean titer
A A+H B B+H A+B+H H
1 18 1 3 3 23 49
volume in developed read after the details
0.82 x 3.34x 0.64 x 4.64x 3.07x 1.7 x
104 104 102 104 10:~ 10~
. Control subjects No. I Mean titer
2 14 3 12
1.64x 1.4x 2.73 x 1.61 x
104 104 10* 102
31
the tubes was 0.5 ml. T h e patterns at the bottom of the tubes were 2 hours a n d 4 hours. T a b l e I lists of the procedure.
RESULTS
T a b l e I I lists the frequencies of secretors a n d nonsecretors. Percentages for patients with cystic fibrosis a n d for control subjects are essentially the same. These frequencies did n o t differ from those reported for the n o r m a l population: 76 to 78 per cent secretors and 22 to 24 per cent nonsecretors. 2, 3 I n addition, no significant difference was f o u n d between titers of the group-specific
Volume 68
Number 1
substances in saliva studied ( T a b l e I I I ) .
Brief clinical and laboratory observations
of
the
two
groups
SUMMARY T h e salivary secretion of A B H blood group substances was studied in 65 patients with cystic fibrosis a n d 40 n o r m a l control subjects. No association was d e m o n s t r a t e d between A B H secretory status and cystic fibrosis.
14 l
REFERENCES 1. Chernick, W. S., and Barbero, G. J.: Studies on human tracheobronchial and submaxillary secretions in normal and pathophysiological conditions, Ann. New York Acad. Sc. 106: 698, 1963. 2. Race, R. R., and Sanger, R.: Blood groups in man, Oxford, 1962, Blackwell Scientific Publications. 3. Wiener, A. S.: Blood groups and transfusion, New York, 1962, Hafner Publishing Company, Inc.
A modified electrode and lead assembly for rectorcardiograpby in newborn and small infants Lorin E. Ainger, M.D., "x"a n d Bill R. C a m p b e l l MEMPHIS,
TENN.
T w o o v the most difficult technical problems of e l e c t r o c a r d i o g r a p h y a n d vectorcard i o g r a p h y in n e w b o r n a n d small infants are m a i n t e n a n c e of firm sldn-to-electrode contact a n d m a i n t e n a n c e of secure lead-electrode connections. A disposable adhesive electrode with snap c o n n e c t o r t p r o v i d e d a satisfacto W solution to these problems. This electrode is a 5//s inch square, silver impregn a t e d meshed cloth fixed to a 11~ inch square plastic adhesive b a n d a g e with an o r d i n a r y g r i p - s n a p p e r to w h i c h the lead is securely fastened. T h e cost of this electrode, however, p r e c l u d e d its use on a large scale; therefore, From the section o/Pediatric Cardiolog~, Department of Pediatrics, University of Tennessee College of Medicine. This work was supported by Grant No. HE-5755-C3, National Heart Institute, National Institutes of Health, United States Public Health Service. "X'Address, 860 Madison Avenue, Memphis, Tenn. 38103. tTelemedies, Inc., Southampton, Pa.
Fig. 1. Components of the modified electrode and lead assembly. From left to right: (1) the electrode, 5/8" in diameter; (2) the 1 ~ " square plastic adhesive bandage with the hole punched in the center; (3) the lead with the connector to fit the grip-snapper.