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Myocardial infarction Changes in the concentrations of high-energy compounds and free amino acids in erythrocytes
359
Atherosclerosis,
40 (1981)
Elsevier/North-Holland
Preliminary
359-364 Scientific Publishers,
Note
MYOCARDIAL
INFARCTION
Changes in the Concentrations Acids in Erythrocytes
of High-energy Compounds
H. K DZIORA, A. LAO, A. GOLIfiSKI, W. T f ACZEWSKI
D. CIESLIfiSKA,
Department of Physiology and Department Medicine, Warsaw (Poland) (Received (Accepted
Ltd.
and Free Amino
J. KFDZIORA
and
of Internal Medicine, Warsaw Academy of
22 April, 1981) 23 April, 1981)
Summary
Quantitative determination of the nucleotides AMP, ADP, ATP, GTP, NAD, NADP, 2,3-DPG and the free amino acids Lys, His, Gly, Ala, Val, Met, Phe, Tyr, Pro, Thr, Ser, Glu, Asp in erythrocytes was carried out in early and late stages of myocardial infarction. It was found that in erythrocytes, in the early stage of myocardial infarction, the concentrations of AMP, NADP and 2,3-DPG increased, whereas those of ADP, ATP, GTP and NAD decreased. In the third week of the disease the concentrations of AMP, ADP, NADP, and especially 2,3-DPG remained high, while those of ATP and GTP shifted towards the control. The concentrations of His, Gly, Ala, Val, Met, Phe, Thr and Glu increased, while those of Tyr, Ser and Asp decreased in the first stage of myocardial infarction. At the later stage of the illness (21 days) the concentrations of free amino acids returned to normal. Key words:
Free amino acids -High-energy Myocardial infarction
compounds -Metabolism
of erythrocytes
-
Introduction
Almost all cells and tissues of the organism are involved in the complex biochemical disturbances that occur during myocardial infarction (MI). Investigation in recent years has.elucidated the action of the adrenergic system on the metabolic pathways of carbohydrates, lipids &d protein changes, and revealed an increase in glycolysis and free fatty acids, increased activity in some of the enzymatic proteins characteristic of MI, and profile changes in the plasma OOZl-9150/81/0000-0000/!$02.50
@ 1981 Elsevier/North-Holland
Scientific
Publishers,
Ltd.
360
structural proteins [l-7]. In spite of these relatively well-described biochemical disturbances, very little is known about the changes occurring in the erythrocytes, which play the most important role in the process of oxygen delivery to the tissues. The aim of this work was to analyse metabolic changes in the erythrocytes with special regard to high-energy compounds and free amino acids. Materials and Methods The investigation was carried out on 29 patients aged 40-72 (mean age: 58) years. The MI was diagnosed by means of generally accepted criteria; anamneses, objective examinations, electrocardiographic changes, and typical routine biochemical indicators such as CPK, Asp-At, Ala-At, glucose, proteinograms, pH, gasometric parameters etc.. The patients were all kept on the same protective diet and given similar analgesic drugs, as well as drugs facilitating coronary circulation. The determination of high energy compounds in erythrocytes (A) was performed in three stages: I, 2-3rd; II, 7-&h; and III, 20-21st days after the beginning of the MI. The evaluation of amino acids in erythrocytes (B) was done on the 2-3rd day and the 20-21st day. Blood (10 ml) was taken in the morning from the cubitus vein, heparinized, then centrifuged for 15 min at about 4 000 rpm at 4°C. The leukocyte pellet was mechanically removed from the erythrocytes, which were then washed three times with 0.9% NaCl solution. (A) The acid-soluble fraction of erythrocytes containing phosphate esters was obtained by Bartlett’s method [ 81 and separated on ion exchange resin according to Hurlbert et al. [9]. (B) The extract of amino acids was obtained according to Kedziora et al. [lo] and determined by ion exchange on a JEOL ILC-64 H amino acid analyser [ 111. Results The following fractions were obtained from the acid-soluble extract of erythrocytes: adenosine 5’-monophosphate (AMP), adenosine 5’-diphosphate (ADP), adenosine 5’-triphosphate (ATP), guanosine 5’-triphosphate (GTP), nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), 2,3-diphosphoglycerate (2,3-DPG). Table 1 shows the concentrations of phosphate esters in erythrocytes of patients with MI at given stages, in comparison with the control. Table 2 lists the amino acid concentrations in haemolysates of erythrocytes in patients with MI when compared with the control. The following amino acids were determined quantitatively: lysine (Lys), histidine (His), glycine (Gly), alanine (Ala), valine (Val), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), proline (Pro), threonine (Thr), serine (Ser), glutamic acid (Glu), and aspartic acid (Asp). The results were subject to statistical analysis using the S-test (0.01 < P d 0.05). The investigation of the content of phosphate esters in erythrocytes suggests that the concentrations of AMP, NADP and 2,3-DPG increase and those of ADP, ATP, GTP and NAD decrease in the first 3 days after the beginning of MI. Assessment of the above-mentioned compounds at the 2nd and 3rd stages of the illness show that the concentrations of AMP, ADP, NADP and especially
361 TABLE 1 COMPARISON OF PHOSPHATE ESTER CONTENT IN ERYTHROCYTES CARDIAL INFARCTION AND THAT IN THE CONTROL GROUP
OF PATIENTS WITH MYO-
Values expressed as fiM/lOO ml erythrocytes. Patients with myocardial infarction
Subject examined
Control group
Duration of examLnation Early stage (2-3 days)
Intermediate stage (7-8 days)
Late stage (20-21 days)
AMP
x SD n
9.970 1.770 12
s
7.338 3.209 11
N
8.628 4.006 10
N
6.866 1.350 10
ADP
x SD n
33.226 10.767 12
N
32.539 11.620 11
N
30.796 4.116 10
s
38.900 4.996 10
ATP
x SD n
72.791 8.634 12
s
109.456 32.009 11
100.312 10.946 10
S
136.130 21.770 10
GTP
x SD n
4.478 1.292 12
S
6.216 1.342 11
N
6.941 1.021 10
N
6.718 1.076 10
NAD
I SD n
3.668 0.469 12
N
4.113 0.407 11
N
4.669 0.861 10
N
4.013 0.761 10
NADP
I SD n
10.784 1.738 12
S
8.918 1.011 11
S
8.926 0.963 10
S
7.533 0.990 10
2,3-DPG
x SD n
416.331 69.367 12
S
441.093 60,467 11
s
602.464 73.646 10
S
367.790 38.644 10
s
X= mean, SD = standard deviation, n = number, S = significant, N = non significant.
2,3-DPG were still maintained at the high level, whereas ATP and GTP were normal (Table 1). At the early stage of MI the concentrations of His, Gly, Ala, Val, Met, Phe, Thr and Glu increased and those of Tyr, Ser, Asp decreased. The concentrations of Pro and Lys did not show any changes. It is worth noting that in the 3rd week of the illness the free amino acids concentration in erythrocytes (Table 2) was approaching normal. Discussion The changes in the concentrations of high-energy compounds in erythrocytes and especially ATP and 2,3-DPG immediately affect the function of these cells, which in turn leads to change8 in the affinity of haemoglobin for oxygen [ 121. 2,3-DPG preferentially binds to deoxygenated haemoglobin, but such binding diminishes haemoglobin oxygen affinity and facilitates oxygen release to tissues, shifting the oxygen dissociation curve of haemoglobin to the right. The high concentration of 2,3-DPG in erythrocytes observed during our experiments at constant pH, pOz and pCOz would suggest the existence of a com-
362 TABLE 2 COMPARISON OF THE AVERAGE VALUE OF FREE AMINO ACIDS IN ERYTHROCYTES PATIENTS WITH MYOCARDIAL INFARCTION AND THAT IN THE CONTROL GROUP Values are expressed in pM/lOO ml of erythrocytes. Amino acid
Patients with myocardial infarction
Control group
Duration of examination Early stage (2-3 days)
x
Late stage (20-21 days)
3.793 0.326 17
N
3.716 0.416 17
N
SD 11
3.649 0.265 10
His
x SD n
1.666 1.080 17
S
1.174 0.170 17
N
1.013 0.100 10
Gly
x SD n
16.468 2.639 17
S
13.397 1.239 17
s
12.133 0.890 10
Ala
x SD n
20.636 1.300 17
S
16.104 0.688 17
S
16.860 0.690 10
Val
x SD n
1.811 0.082 17
1.369 0.136 17
N
1.477 0.114 10
Met
x SD 11
0.382 0.047 17
S
0.238 0.029 16
N
0.276 0.047 10
Phe
y SD n
0.903 0.047 17
N
0.891 0.061 17
N
0.872 0.086 10
Tyr
x SD n
1.319 0.082 17
s
2.060 0.228 17
S
1.747 0.146 10
pro
x
1.032 0.206 17
N
0.864 0.109 17
N
0.930 0.146 10
Thr
SD n x SD n
3.082 0.269 17
8
2.875 0.207 17
N
2.860 0.225 10
x SD n
3.278 0.436 17
8
4.122 0.371 17
N
4.772
Glu
r SD n
14.088 3.661 17
8
9.350 1.739 17
8
7.710 0.861 10
A.sP
x SD n
12.069 1.894 17
s
14.416 0.678 17
S
18.466 0.666 10
LYS
Se:
S
0.284 10
E= mean. SD - standard devation. n = number, 8 = significmt. N = non significant.
OF
363
pensating mechanism in hypoxic tissues that require large oxygen supplies [ 13, 141. The red cell is one of the models in which active amino acid transport has been examined, in both the physiological and pathological states [ 15-171. Our findings indicate that in the early stages of MI the concentration of the majority of the amino acids increases both in the plasma [ 181 and the erythrocytes. This can be explained by the predominance of catabolic processes in early stage of the illness. The data we obtained are in agreement with previously published work [ 19-211. It is interesting to note that in the early stage of the illness the concentration of Glu increases in the erythrocytes; Krzeczkowska et al. [22] have observed this increase of glutamic acid in plasma. The low concentration of aspartic acid in erythrocytes seems to us to be worthy of note. It may be due to the intensification of transamination processes caused by the increase of amino transferase activity. Acknowledgements We are grateful to Professor G.V.R. Born, F.R.C.P., F.R.S., London King’s College, England, for many helpful discussions.
University of
References 1 Berg, K., Dehlen, G. and Frik. M.H., Lp (a) Lipoprotein and pre-&-lipoprotein in patients with corenary heart dieeene. Clin. Genet., 6 (1974) 230. 2 Jeguier, E. and Perret. C.. Urinary excretion of catecholeminee and their main metabolites after myocardial infarction, Europ. J. Clin. Invest., 1 (1970) 77. 3 Opie. L.H.. Metabolism of free fatty acide. glucose end catecholamines in acute myocardial fnfarction -Relation to myocardial iechemia and infarct dze, Amer. J. Caniiol., 36 (1976) 938. 4 Hearse, D.J., Myocardial enzyme leakage, J. Mol. Med., 2 (1977) 185. 6 Kekki, M. and Helonen. P.J.. Studies of plasma and urinary amino nitrogen in myocardial infarction. Acta Med. Stand., 168 (1960) 133. 6 Kahn, J.C., Gueret. P., Menier, R.. Girendet. P.. Farhat. M.B. and Bourdariae. J.P., Prognostic velue of enzymatic (CPK) estimation of infarct dze. J. Mol. Med.. 2 (1977) 233. 7 Me&r, H.S., Evans, R. and Ayers, S.M.. Effect of dopamine on hemodynamics and myocerdiel metaboliem in chock following acute myocardial infarction in man, Circulation. 87 (1978) 361. 8 Bartlett, G.R.. Methods for isolation of glycolytic intermediates by column chromatography with ion exchange redns, J. Biol. Chem.. 234 (1959) 469. 9 Hurlbert, R.B.. Schmitz. H., Brum, A.F. and Potter, V.R.. Nucleotide metabolism, Part 2 (Chromatographic separation of acid soluble nucleotider, J. Biol. Chem., 209 (1964) 23. 10 Kpdziora, J.. Kpdziora. H. and Jeeke. J.. Free amino acids in erythrocytes of children with Down’s syndrome. Endokr. Pal.. 24 (1973) 219 (in Polish). 11 Stein, W.H. and Moore, S.. The amino acids of human blood plasma, J. Biol. Chem.. 211 (1964) 916. 12 Baer, 5.. Anaemla of bums, Burne, 6 (1979) 1. 13 Paniker. N.V. and Beutler. E.. Effecb of normal metabolites on the oxygen-haemoglobin equilibrium. Rot. Sot. Exp. Biol. Med., 136 (1970) 389. 14 Surgenor, D.M., Traneport of oxygen and carbon dioxide, In: Ch. Biabop and D.M. Surgenor (Ed%). The Red Blood Cell, Academic Pren. New York, 1964. p. 346. 16 Nekao, M. and Nakao. T.. Biochemietry of the erythrocyte membrane with emphadB on ene4Y metabolbm. Adan Med. J.. 19 (1976) 7. 16 Nekao, M. and Nakayama. T., Decreeee in phoephofructokinaee activity during blood preservation and the effect of intracelluLr ATP. Biochem. Biophys. Rec. Comm., 95 (1980) 1294. 17 Bishop. Ch.. protein end imino acid metaboB.em. In: Ch. Btehop and D.M. Surgenor (Ed%). The Red Blood Cell, Academic Press. New York. 1964. p. 178. 18 K@iora, H.. Conaentration of amino acide in plasma and erythrocytee in patients with myowdw infezction, Kard. Pol.. 23 (1980) 299 (in Polish).
19 Efimova, L.G., Ch~nper in the serum content of some free amino acids during acute myocesdial inferction. Cor et Vaea. 6 (1963) 30. 20 Widomeka-Czajkoweka, T.. Aminoacidaemiae in myocardial infarction, Pol. Tyg. Lek., 23 (1969) 1637 (in Polish). 21 D@broweki. A. and Iwa6eka. J.. Changes in amino acids concentration in serum during myocardial infarction. Pol. Arch. Med. Wewn.. 52 (1974) 149 (in Pouch). 22 Kneczkowaka. J., Czexniek, Z. and Burzyfiski. S.. Quantitative determination of free amino acids in blood of patiente with myocardiel infarction. Pol. Arch. Med. Wewn.. 41(1966) 361 (in Polieh).
Atherosclerosis,
40 (1981)
Elsevier/North-Holland
Preliminary
359-364 Scientific Publishers,
Note
MYOCARDIAL
INFARCTION
Changes in the Concentrations Acids in Erythrocytes
of High-energy Compounds
H. K DZIORA, A. LAO, A. GOLIfiSKI, W. T f ACZEWSKI
D. CIESLIfiSKA,
Department of Physiology and Department Medicine, Warsaw (Poland) (Received (Accepted
Ltd.
and Free Amino
J. KFDZIORA
and
of Internal Medicine, Warsaw Academy of
22 April, 1981) 23 April, 1981)
Summary
Quantitative determination of the nucleotides AMP, ADP, ATP, GTP, NAD, NADP, 2,3-DPG and the free amino acids Lys, His, Gly, Ala, Val, Met, Phe, Tyr, Pro, Thr, Ser, Glu, Asp in erythrocytes was carried out in early and late stages of myocardial infarction. It was found that in erythrocytes, in the early stage of myocardial infarction, the concentrations of AMP, NADP and 2,3-DPG increased, whereas those of ADP, ATP, GTP and NAD decreased. In the third week of the disease the concentrations of AMP, ADP, NADP, and especially 2,3-DPG remained high, while those of ATP and GTP shifted towards the control. The concentrations of His, Gly, Ala, Val, Met, Phe, Thr and Glu increased, while those of Tyr, Ser and Asp decreased in the first stage of myocardial infarction. At the later stage of the illness (21 days) the concentrations of free amino acids returned to normal. Key words:
Free amino acids -High-energy Myocardial infarction
compounds -Metabolism
of erythrocytes
-
Introduction
Almost all cells and tissues of the organism are involved in the complex biochemical disturbances that occur during myocardial infarction (MI). Investigation in recent years has.elucidated the action of the adrenergic system on the metabolic pathways of carbohydrates, lipids &d protein changes, and revealed an increase in glycolysis and free fatty acids, increased activity in some of the enzymatic proteins characteristic of MI, and profile changes in the plasma OOZl-9150/81/0000-0000/!$02.50
@ 1981 Elsevier/North-Holland
Scientific
Publishers,
Ltd.
360
structural proteins [l-7]. In spite of these relatively well-described biochemical disturbances, very little is known about the changes occurring in the erythrocytes, which play the most important role in the process of oxygen delivery to the tissues. The aim of this work was to analyse metabolic changes in the erythrocytes with special regard to high-energy compounds and free amino acids. Materials and Methods The investigation was carried out on 29 patients aged 40-72 (mean age: 58) years. The MI was diagnosed by means of generally accepted criteria; anamneses, objective examinations, electrocardiographic changes, and typical routine biochemical indicators such as CPK, Asp-At, Ala-At, glucose, proteinograms, pH, gasometric parameters etc.. The patients were all kept on the same protective diet and given similar analgesic drugs, as well as drugs facilitating coronary circulation. The determination of high energy compounds in erythrocytes (A) was performed in three stages: I, 2-3rd; II, 7-&h; and III, 20-21st days after the beginning of the MI. The evaluation of amino acids in erythrocytes (B) was done on the 2-3rd day and the 20-21st day. Blood (10 ml) was taken in the morning from the cubitus vein, heparinized, then centrifuged for 15 min at about 4 000 rpm at 4°C. The leukocyte pellet was mechanically removed from the erythrocytes, which were then washed three times with 0.9% NaCl solution. (A) The acid-soluble fraction of erythrocytes containing phosphate esters was obtained by Bartlett’s method [ 81 and separated on ion exchange resin according to Hurlbert et al. [9]. (B) The extract of amino acids was obtained according to Kedziora et al. [lo] and determined by ion exchange on a JEOL ILC-64 H amino acid analyser [ 111. Results The following fractions were obtained from the acid-soluble extract of erythrocytes: adenosine 5’-monophosphate (AMP), adenosine 5’-diphosphate (ADP), adenosine 5’-triphosphate (ATP), guanosine 5’-triphosphate (GTP), nicotinamide adenine dinucleotide (NAD), nicotinamide adenine dinucleotide phosphate (NADP), 2,3-diphosphoglycerate (2,3-DPG). Table 1 shows the concentrations of phosphate esters in erythrocytes of patients with MI at given stages, in comparison with the control. Table 2 lists the amino acid concentrations in haemolysates of erythrocytes in patients with MI when compared with the control. The following amino acids were determined quantitatively: lysine (Lys), histidine (His), glycine (Gly), alanine (Ala), valine (Val), methionine (Met), phenylalanine (Phe), tyrosine (Tyr), proline (Pro), threonine (Thr), serine (Ser), glutamic acid (Glu), and aspartic acid (Asp). The results were subject to statistical analysis using the S-test (0.01 < P d 0.05). The investigation of the content of phosphate esters in erythrocytes suggests that the concentrations of AMP, NADP and 2,3-DPG increase and those of ADP, ATP, GTP and NAD decrease in the first 3 days after the beginning of MI. Assessment of the above-mentioned compounds at the 2nd and 3rd stages of the illness show that the concentrations of AMP, ADP, NADP and especially
361 TABLE 1 COMPARISON OF PHOSPHATE ESTER CONTENT IN ERYTHROCYTES CARDIAL INFARCTION AND THAT IN THE CONTROL GROUP
OF PATIENTS WITH MYO-
Values expressed as fiM/lOO ml erythrocytes. Patients with myocardial infarction
Subject examined
Control group
Duration of examLnation Early stage (2-3 days)
Intermediate stage (7-8 days)
Late stage (20-21 days)
AMP
x SD n
9.970 1.770 12
s
7.338 3.209 11
N
8.628 4.006 10
N
6.866 1.350 10
ADP
x SD n
33.226 10.767 12
N
32.539 11.620 11
N
30.796 4.116 10
s
38.900 4.996 10
ATP
x SD n
72.791 8.634 12
s
109.456 32.009 11
100.312 10.946 10
S
136.130 21.770 10
GTP
x SD n
4.478 1.292 12
S
6.216 1.342 11
N
6.941 1.021 10
N
6.718 1.076 10
NAD
I SD n
3.668 0.469 12
N
4.113 0.407 11
N
4.669 0.861 10
N
4.013 0.761 10
NADP
I SD n
10.784 1.738 12
S
8.918 1.011 11
S
8.926 0.963 10
S
7.533 0.990 10
2,3-DPG
x SD n
416.331 69.367 12
S
441.093 60,467 11
s
602.464 73.646 10
S
367.790 38.644 10
s
X= mean, SD = standard deviation, n = number, S = significant, N = non significant.
2,3-DPG were still maintained at the high level, whereas ATP and GTP were normal (Table 1). At the early stage of MI the concentrations of His, Gly, Ala, Val, Met, Phe, Thr and Glu increased and those of Tyr, Ser, Asp decreased. The concentrations of Pro and Lys did not show any changes. It is worth noting that in the 3rd week of the illness the free amino acids concentration in erythrocytes (Table 2) was approaching normal. Discussion The changes in the concentrations of high-energy compounds in erythrocytes and especially ATP and 2,3-DPG immediately affect the function of these cells, which in turn leads to change8 in the affinity of haemoglobin for oxygen [ 121. 2,3-DPG preferentially binds to deoxygenated haemoglobin, but such binding diminishes haemoglobin oxygen affinity and facilitates oxygen release to tissues, shifting the oxygen dissociation curve of haemoglobin to the right. The high concentration of 2,3-DPG in erythrocytes observed during our experiments at constant pH, pOz and pCOz would suggest the existence of a com-
362 TABLE 2 COMPARISON OF THE AVERAGE VALUE OF FREE AMINO ACIDS IN ERYTHROCYTES PATIENTS WITH MYOCARDIAL INFARCTION AND THAT IN THE CONTROL GROUP Values are expressed in pM/lOO ml of erythrocytes. Amino acid
Patients with myocardial infarction
Control group
Duration of examination Early stage (2-3 days)
x
Late stage (20-21 days)
3.793 0.326 17
N
3.716 0.416 17
N
SD 11
3.649 0.265 10
His
x SD n
1.666 1.080 17
S
1.174 0.170 17
N
1.013 0.100 10
Gly
x SD n
16.468 2.639 17
S
13.397 1.239 17
s
12.133 0.890 10
Ala
x SD n
20.636 1.300 17
S
16.104 0.688 17
S
16.860 0.690 10
Val
x SD n
1.811 0.082 17
1.369 0.136 17
N
1.477 0.114 10
Met
x SD 11
0.382 0.047 17
S
0.238 0.029 16
N
0.276 0.047 10
Phe
y SD n
0.903 0.047 17
N
0.891 0.061 17
N
0.872 0.086 10
Tyr
x SD n
1.319 0.082 17
s
2.060 0.228 17
S
1.747 0.146 10
pro
x
1.032 0.206 17
N
0.864 0.109 17
N
0.930 0.146 10
Thr
SD n x SD n
3.082 0.269 17
8
2.875 0.207 17
N
2.860 0.225 10
x SD n
3.278 0.436 17
8
4.122 0.371 17
N
4.772
Glu
r SD n
14.088 3.661 17
8
9.350 1.739 17
8
7.710 0.861 10
A.sP
x SD n
12.069 1.894 17
s
14.416 0.678 17
S
18.466 0.666 10
LYS
Se:
S
0.284 10
E= mean. SD - standard devation. n = number, 8 = significmt. N = non significant.
OF
363
pensating mechanism in hypoxic tissues that require large oxygen supplies [ 13, 141. The red cell is one of the models in which active amino acid transport has been examined, in both the physiological and pathological states [ 15-171. Our findings indicate that in the early stages of MI the concentration of the majority of the amino acids increases both in the plasma [ 181 and the erythrocytes. This can be explained by the predominance of catabolic processes in early stage of the illness. The data we obtained are in agreement with previously published work [ 19-211. It is interesting to note that in the early stage of the illness the concentration of Glu increases in the erythrocytes; Krzeczkowska et al. [22] have observed this increase of glutamic acid in plasma. The low concentration of aspartic acid in erythrocytes seems to us to be worthy of note. It may be due to the intensification of transamination processes caused by the increase of amino transferase activity. Acknowledgements We are grateful to Professor G.V.R. Born, F.R.C.P., F.R.S., London King’s College, England, for many helpful discussions.
University of
References 1 Berg, K., Dehlen, G. and Frik. M.H., Lp (a) Lipoprotein and pre-&-lipoprotein in patients with corenary heart dieeene. Clin. Genet., 6 (1974) 230. 2 Jeguier, E. and Perret. C.. Urinary excretion of catecholeminee and their main metabolites after myocardial infarction, Europ. J. Clin. Invest., 1 (1970) 77. 3 Opie. L.H.. Metabolism of free fatty acide. glucose end catecholamines in acute myocardial fnfarction -Relation to myocardial iechemia and infarct dze, Amer. J. Caniiol., 36 (1976) 938. 4 Hearse, D.J., Myocardial enzyme leakage, J. Mol. Med., 2 (1977) 185. 6 Kekki, M. and Helonen. P.J.. Studies of plasma and urinary amino nitrogen in myocardial infarction. Acta Med. Stand., 168 (1960) 133. 6 Kahn, J.C., Gueret. P., Menier, R.. Girendet. P.. Farhat. M.B. and Bourdariae. J.P., Prognostic velue of enzymatic (CPK) estimation of infarct dze. J. Mol. Med.. 2 (1977) 233. 7 Me&r, H.S., Evans, R. and Ayers, S.M.. Effect of dopamine on hemodynamics and myocerdiel metaboliem in chock following acute myocardial infarction in man, Circulation. 87 (1978) 361. 8 Bartlett, G.R.. Methods for isolation of glycolytic intermediates by column chromatography with ion exchange redns, J. Biol. Chem.. 234 (1959) 469. 9 Hurlbert, R.B.. Schmitz. H., Brum, A.F. and Potter, V.R.. Nucleotide metabolism, Part 2 (Chromatographic separation of acid soluble nucleotider, J. Biol. Chem., 209 (1964) 23. 10 Kpdziora, J.. Kpdziora. H. and Jeeke. J.. Free amino acids in erythrocytes of children with Down’s syndrome. Endokr. Pal.. 24 (1973) 219 (in Polish). 11 Stein, W.H. and Moore, S.. The amino acids of human blood plasma, J. Biol. Chem.. 211 (1964) 916. 12 Baer, 5.. Anaemla of bums, Burne, 6 (1979) 1. 13 Paniker. N.V. and Beutler. E.. Effecb of normal metabolites on the oxygen-haemoglobin equilibrium. Rot. Sot. Exp. Biol. Med., 136 (1970) 389. 14 Surgenor, D.M., Traneport of oxygen and carbon dioxide, In: Ch. Biabop and D.M. Surgenor (Ed%). The Red Blood Cell, Academic Pren. New York, 1964. p. 346. 16 Nekao, M. and Nakao. T.. Biochemietry of the erythrocyte membrane with emphadB on ene4Y metabolbm. Adan Med. J.. 19 (1976) 7. 16 Nekao, M. and Nakayama. T., Decreeee in phoephofructokinaee activity during blood preservation and the effect of intracelluLr ATP. Biochem. Biophys. Rec. Comm., 95 (1980) 1294. 17 Bishop. Ch.. protein end imino acid metaboB.em. In: Ch. Btehop and D.M. Surgenor (Ed%). The Red Blood Cell, Academic Press. New York. 1964. p. 178. 18 K@iora, H.. Conaentration of amino acide in plasma and erythrocytee in patients with myowdw infezction, Kard. Pol.. 23 (1980) 299 (in Polish).
19 Efimova, L.G., Ch~nper in the serum content of some free amino acids during acute myocesdial inferction. Cor et Vaea. 6 (1963) 30. 20 Widomeka-Czajkoweka, T.. Aminoacidaemiae in myocardial infarction, Pol. Tyg. Lek., 23 (1969) 1637 (in Polish). 21 D@broweki. A. and Iwa6eka. J.. Changes in amino acids concentration in serum during myocardial infarction. Pol. Arch. Med. Wewn.. 52 (1974) 149 (in Pouch). 22 Kneczkowaka. J., Czexniek, Z. and Burzyfiski. S.. Quantitative determination of free amino acids in blood of patiente with myocardiel infarction. Pol. Arch. Med. Wewn.. 41(1966) 361 (in Polieh).