EI SOON CHO,6 GARY F. KRAUSE ANDHELEN L. ANDERSON Department of Human Nutrition, Foods and Food Systems Management, and Department of Statistics and Agricultural Experiment Station, University of Missouri, Columbia, Missouri 65201 ABSTRACT The effects of dietary histidine and arginine on fasting and 1 and 2 hour postprandial plasma free amino acid and urea concentrations were studied in six young men. For 1 week each, they were fed six differ ent diets containing 6.3 g of nitrogen daily. Each diet contained eight indispensable amino acids, cystine and tyrosine proportioned as in casein and a different mixture of dispensable nitrogen: A) six dispensable amino acids plus arginine (diet 1) or plus histidine and arginine (diet 2) in the casein pattern, or B ) an isonitrogenous amount of glycine and diammonium citrate alone (diet 3), with histidine (diet 4), with arginine (diet 5) or with histidine and arginine (diet 6). The fasting plasma concentrations of the seven indispensable amino acids assayed and their similar postprandial patterns were unaffected by the dietary treatments. Both fasting and post prandial plasma histidine concentrations were significantly lower when the histidine-low diets were fed than when the histidine-supplemented diets were fed. Histidine supplementation promoted a reduction in fasting plasma urea nitrogens. Proline concentrations were lowered significantly when proline was removed from the dietary amino acid mixtures, but plasma arginine concentrations were unaffected by arginine removal. Plasma histi dine was maintained at lower concentrations in dietary histidine deficiency than when histidine was added to the low nitrogen diets. J. Nutr. 107: 2078-2089, 1977. INDEXING KEY WORDS histidine •arginine •plasma amino acids •plasma urea •adult humans A significant amount of research has been directed toward the correlation of plasma amino acid concentrations with amino acid adequacy of the test diet (1-10).
amino
Although
acid
monitoring
pattern
may
the
not
i . i r enough concrete evidence for marcnnal aminn aHrl r)f>finif>nrioc marginai amino acia Deficiencies eral. investigators i T?. . have i successfully ... .
mined arid
qualitative requirements
USing changes in Centrations either rentTT- nitrOSen I-i-
Misname '
plasma
plasma amino with Or without
balance fiance
data aata
Received for publication
November
8, 1976.
provide
»*J,°e,rnal ,pape/ JJ0-,7695 of the Missouri Agricui-
-i detecting (Q\ (»;, «nr sev' deter-.
turai Experiment Station. »Part of a dissertation submitted by El Soon Cho to University for of Missouri partial fulfillment of the tne requirements the Ph.D.in degree. , 'Presented of in Institute Nutrition, part at the Atlantic meetingCity, of the NewAmerican Jersey,
and quantitative ammo in animale anrl ninnane in animais ana numans
rpmiirpmpnr«
acids for the maintenance of nitrogen balance in adult males (18-20). In studies of amino acid requirements, Rose obtained
acid COnconcur-
110-17 \ (.IV-U}.
and arginine have been classioo /J^Li-r.», JUl • as dietary dispensable amino
April, 1974; Federation Proc. 33, 711 (Abstr.). « Supported in part by National Institutes of Health Research Grant NO. AMISSSS,The Nutrition Foundation, Inc. Grant No. 450, and the Missouri AgrlCUsThe rerearch^protocol "received approval from the Committee for Research Involving Human Subjects, School of Medicine, University of Missouri-Columbia
prior to initiation of the experiment. «Present address: Department of Home Economics, university of iowa, iowa city, Iowa 52242. 2078
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
Effects of dietary histidine and arginine on plasma amino acid and urea concentrations of men fed a low nitrogen diet1-s
HISTIDINE, ARGININE AND PLASMA AMINO ACIDS
EXPERIMENTAL PROCEDURES
The diets, vital statistics of the subjects, and most of the experimental procedures have been reported in detail in the previous paper (25). Experimental design. The 57-day experi ment consisted of a 3-day depletion period, a 5-day adjustment period, and seven 7-day experimental periods. The experimental
TABLE 1 Daily amount of amino acids and diammonium citrate furnished by the amino acid mixtures in each experimental diet1-* numberAmino acid12»3»4>
Diet 5'6*O/
dayLsubject/ ValineL-IsoleucineL-LeucineL-ThreonineL-MethionineL-PhenylalanineL-Lysine
HC1L-TryptophanL-CyatineL-TyrosineL-AlanineGlycineL-ProlineL-SerineL-Aspartic •
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
positive nitrogen retention in over 50 men ted diets devoid of these two amino acids (21). The incorporation of 15NH4C1 into the imidazole ring indicated the synthesis of histidine by one adult male receiving histidine-free total parenteral nutrition (22). Nevertheless, beneficial effects of histidine and arginine on nitrogen balance (23-25) have been reported. Elevated serum activities of glutamic-pyruvic and glutamic-oxalacetic transaminase reported in men fed diets deficient in arginine and histidine (26) have not been substantiated (24, 25, 27). Also, decreases have been reported in fasting and/or postprandial plasma histidine (28-30), or histidine and arginine ( 31 ), concentrations of men when these two amino acids were not included in the dietary amino acid mixtures provid ing a large proportion of the total daily nitrogen. A trend toward reduced plasma histidine concentration in the near absence of dietary histidine was observed in the study of Weller et al. (32), although the decrease was not statistically significant. In addition, it has been suggested that histi dine and/or arginine, most probably his tidine, may be an indispensable amino acid for young men (26, 30, 33). In view of these findings, a study was conducted to specifically test for the ef fects of dietary histidine and arginine on plasma amino acid concentrations and ni trogen utilization. Previously reported data (25) from this study showed that nitrogen balance was significantly improved when young men consumed a histidine-supplemented crystalline amino acid diet com pared to a histidine-low diet, regardless of the other sources of dispensable nitrogen provided. The present paper reports the effects of dietary histidine and arginine on the concentrations of individual free amino acids and urea in the plasma of these same young men.
2079
07.70 7.700 00 00 acidL-Glutaraic 00 acidL00 Arginine•HC1L-Histidine•HC1-H.ODiammoniumcitrate2.562.263.481.482.071.863.460.500.132.01 1.701.41 013.43 12.0607.7000001.701.419.78 1These amounts do not include the amino acids contained in the 0.81 g N in the ordinary foods of the diet. ! Amino acids are patterned as in casein, with the exception of glycine and diammonium citrate in diets 3 through 6, and with additional methionine and tryptophan to meet twice the minimum re quirements of Rose (21). ' Levels of each indispensable amino acid plus cystine and tyrosine in all diets were the same as in diet 1.
design (25) was a 6x6 extra-period, Latin-square, change-over design balanced for one-period residual effects (34). The diet consumed by each subject in the sixth experimental period was also consumed in the seventh period. Each subject was per mitted to receive diets without histidine for no longer than two consecutive 7-day periods. Analysis of variance and least sig nificant differences between means were computed as the test of significance (35). Diets. Crystalline amino acids and di ammonium citrate provided 87% of the daily nitrogen intake of 6.29 g, and the remainder was provided by a few ordinary foods low in nitrogen (25). Daily amounts of amino acids and diammonium citrate furnished by the amino acid mixtures in each of the six experimental diets are given in table 1. The six diets differed in the composition of the dispensable amino acids;
2080
CHO, KRAUSE AND ANDERSON
blood collection, the subjects consumed one-third of the daily allotment of foods within 20 minutes and venous blood was drawn 1 and 2 hours after the beginning numberAmino Diet of the meal. Plasma was obtained from each blood acid123456mg/100 sample by centrifuging at 1,800 X g for 20 minutes at 5°.A portion of the plasma from miValineIsoleucineLancineThreonineMethioninePhenylalanineLysineAlanineGlycine!Proline1SerineAspartic each subject was deproteinized with 1% picric acid, centrifuged, and the picrate removed from the supernatant by ionexchange chromatography; the eluent was analyzed for 16 amino acids by the method of Gehrke et al.7 (36) using a gas chromatograph.8 Transexamic acid (trans-4(aminomethyl)-cyclohexane-carboxylic acid) was used as the internal standard and the acidGlutamic final volatile amino acid derivative pre acidTyrosineHistidine"-'AT'-Methyl-histidine*ArginineTotal pared was the N-trifluoro-acetyl-n-butyl ester. A second portion of plasma was de proteinized with 15% sulfosalicylic acid and centrifuged at 27,000 X g for 15 min utes at 5°.The supernatant was analyzed aminoacidsTotal for lysine, histidine, NT-methylhistidine and indispen A/T-methylhistidine using an ion-exchange aminoacidsTotal sable amino acid analyzer9 according to the dispen method of Speckman et al. (37). Plasma sable aminoacids2.71.42.21.90.41.32.85.62.6«3.0°1.70.88.01.30.7-0.032.137.812.824.02.61.42.12.00.61.23.24.82.6*3.1«2.00.89.31.41.5»0.062.040.713.126.02.81.51.92.20.6 urea nitrogen was determined using the method of Foster and Hohholzer (38). 1These amino acida were determined by the method of Each plasma sample from each subject was Zumwalt et al. (30) using a gas-liquid Chromatograph, except analyzed separately. TABLE 2 Mean fasting plasma amino acid concentrations of men consuming the six experimental diets1•*
abbreviations used to describe these six diets appear in figures 1 and 3. Upon omis sion of histidine or arginine from the amino acid mixtures, diammonium citrate was used to keep the total nitrogen constant. The basal diet was calculated to contain 89 mg of histidine and 175 mg of arginine. Methods. Blood from the antecubital vein of each subject was collected in heparinized tubes before breakfast on the first day of the study, at the end of the adjustment period and on the last day of each 7-day experimental period. Following
RESULTS Fasting plasma amino acid concentra tions. The mean plasma amino acid con centrations of the six subjects before break fast on the seventh day of consuming each of the six experimental diets are presented in table 2. Lysine and histidine concentra tions determined by the ion-exchange or the gas Chromatographie (not shown) methods were approximately the same. The AKmethylhistidine (3-methylhistidine ) con centrations were obtained only from the ion-exchange amino acid analyzer. There fore, the histidine and IVT-methylhistidine values reported are those obtained from the ion-exchange amino acid analyzer, and 7 Gehrke, C. W., Roach, C., Zumwalt, R. W., Stalling, D. L. & Wall, L. L. (1968) Quantitative Gas-Liquid Chromatography of Amino Acids in Proteins and Biological Substances. Analytical Biochemistry Lab oratories, Columbia, Missouri. 8 Bendix model 2500, Bendlx Corporation, Ronceverte, West Virginia. • Beckman model 121, Beckman Instruments, Inc., Splnco Division, Palo Alto, California.
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
for histidine and #r-methylhistidine (see footnote 5). *Means are for six subjects. BEMvalues for the individual amino acida at O, 1, and 2 hr are presented in figures 2 to 4 and table 3. SEM values for the total amino acids, total indispensable and total dispensable amino acids at 0 hr are 2.4, 0.9, and 1.6, respec tively. » Statistically significant differences at P < 0.05 for glycine and proline and at P < 0.01 for histidine are indicated by small letters; diets sharing a common letter are not sig nificantly different. *Diets 1, 3, and 5 versus diets 2, 4, and 6 are significantly different at P < 0.01. 5 Determined by the method of Spackman et al. (37) using an ion-exchange amino acid analyzer.
2081
HISTIDINE, ARGININE AND PLASMA AMINO ACIDS
1 0.66 2 0.42
b1,3, Diets0 hr:
bs
6«1 4.
vs.05
10 l
2| 1 2A*.b2, 1 21••— 1 1 2A«valuation:». ]2His Hours ifter a mael 4Diet 1 Number, +ArgAStatisticalb0 IAA
RM|
'IAA
+
-0 4-- Ì
25+1
26Hit1
DAC —Arg^»,
Diet ]-HitAa0 Competition + Arg3
+ Arghr
Fig. 1 Mean plasma urea nitrogen levels of six young men fed diets differing in source of nonspecific nitrogen. Abbreviations at the bottom of the figure refer to the mixture of eight indispensable amino acids plus cystine and tyrosine (IAA), of six dispensable amino acids (DAA), glycine (Gly), diammonium citrate (DAC), histidine (His), and arginine (Arg). Means are the average values for all six subjects on the last day of each 7-day experimental period in plasma obtained before breakfast (0 hour) and at 1 and 2 hours following breakfast. Small letters indicating statistical evaluations ( P
amino
Although
acid
monitoring
pattern
may
the
not
i . i r enough concrete evidence for marcnnal aminn aHrl r)f>finif>nrioc marginai amino acia Deficiencies eral. investigators i T?. . have i successfully ... .
mined arid
qualitative requirements
USing changes in Centrations either rentTT- nitrOSen I-i-
Misname '
plasma
plasma amino with Or without
balance fiance
data aata
Received for publication
November
8, 1976.
provide
»*J,°e,rnal ,pape/ JJ0-,7695 of the Missouri Agricui-
-i detecting (Q\ (»;, «nr sev' deter-.
turai Experiment Station. »Part of a dissertation submitted by El Soon Cho to University for of Missouri partial fulfillment of the tne requirements the Ph.D.in degree. , 'Presented of in Institute Nutrition, part at the Atlantic meetingCity, of the NewAmerican Jersey,
and quantitative ammo in animale anrl ninnane in animais ana numans
rpmiirpmpnr«
acids for the maintenance of nitrogen balance in adult males (18-20). In studies of amino acid requirements, Rose obtained
acid COnconcur-
110-17 \ (.IV-U}.
and arginine have been classioo /J^Li-r.», JUl • as dietary dispensable amino
April, 1974; Federation Proc. 33, 711 (Abstr.). « Supported in part by National Institutes of Health Research Grant NO. AMISSSS,The Nutrition Foundation, Inc. Grant No. 450, and the Missouri AgrlCUsThe rerearch^protocol "received approval from the Committee for Research Involving Human Subjects, School of Medicine, University of Missouri-Columbia
prior to initiation of the experiment. «Present address: Department of Home Economics, university of iowa, iowa city, Iowa 52242. 2078
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
Effects of dietary histidine and arginine on plasma amino acid and urea concentrations of men fed a low nitrogen diet1-s
HISTIDINE, ARGININE AND PLASMA AMINO ACIDS
EXPERIMENTAL PROCEDURES
The diets, vital statistics of the subjects, and most of the experimental procedures have been reported in detail in the previous paper (25). Experimental design. The 57-day experi ment consisted of a 3-day depletion period, a 5-day adjustment period, and seven 7-day experimental periods. The experimental
TABLE 1 Daily amount of amino acids and diammonium citrate furnished by the amino acid mixtures in each experimental diet1-* numberAmino acid12»3»4>
Diet 5'6*O/
dayLsubject/ ValineL-IsoleucineL-LeucineL-ThreonineL-MethionineL-PhenylalanineL-Lysine
HC1L-TryptophanL-CyatineL-TyrosineL-AlanineGlycineL-ProlineL-SerineL-Aspartic •
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
positive nitrogen retention in over 50 men ted diets devoid of these two amino acids (21). The incorporation of 15NH4C1 into the imidazole ring indicated the synthesis of histidine by one adult male receiving histidine-free total parenteral nutrition (22). Nevertheless, beneficial effects of histidine and arginine on nitrogen balance (23-25) have been reported. Elevated serum activities of glutamic-pyruvic and glutamic-oxalacetic transaminase reported in men fed diets deficient in arginine and histidine (26) have not been substantiated (24, 25, 27). Also, decreases have been reported in fasting and/or postprandial plasma histidine (28-30), or histidine and arginine ( 31 ), concentrations of men when these two amino acids were not included in the dietary amino acid mixtures provid ing a large proportion of the total daily nitrogen. A trend toward reduced plasma histidine concentration in the near absence of dietary histidine was observed in the study of Weller et al. (32), although the decrease was not statistically significant. In addition, it has been suggested that histi dine and/or arginine, most probably his tidine, may be an indispensable amino acid for young men (26, 30, 33). In view of these findings, a study was conducted to specifically test for the ef fects of dietary histidine and arginine on plasma amino acid concentrations and ni trogen utilization. Previously reported data (25) from this study showed that nitrogen balance was significantly improved when young men consumed a histidine-supplemented crystalline amino acid diet com pared to a histidine-low diet, regardless of the other sources of dispensable nitrogen provided. The present paper reports the effects of dietary histidine and arginine on the concentrations of individual free amino acids and urea in the plasma of these same young men.
2079
07.70 7.700 00 00 acidL-Glutaraic 00 acidL00 Arginine•HC1L-Histidine•HC1-H.ODiammoniumcitrate2.562.263.481.482.071.863.460.500.132.01 1.701.41 013.43 12.0607.7000001.701.419.78 1These amounts do not include the amino acids contained in the 0.81 g N in the ordinary foods of the diet. ! Amino acids are patterned as in casein, with the exception of glycine and diammonium citrate in diets 3 through 6, and with additional methionine and tryptophan to meet twice the minimum re quirements of Rose (21). ' Levels of each indispensable amino acid plus cystine and tyrosine in all diets were the same as in diet 1.
design (25) was a 6x6 extra-period, Latin-square, change-over design balanced for one-period residual effects (34). The diet consumed by each subject in the sixth experimental period was also consumed in the seventh period. Each subject was per mitted to receive diets without histidine for no longer than two consecutive 7-day periods. Analysis of variance and least sig nificant differences between means were computed as the test of significance (35). Diets. Crystalline amino acids and di ammonium citrate provided 87% of the daily nitrogen intake of 6.29 g, and the remainder was provided by a few ordinary foods low in nitrogen (25). Daily amounts of amino acids and diammonium citrate furnished by the amino acid mixtures in each of the six experimental diets are given in table 1. The six diets differed in the composition of the dispensable amino acids;
2080
CHO, KRAUSE AND ANDERSON
blood collection, the subjects consumed one-third of the daily allotment of foods within 20 minutes and venous blood was drawn 1 and 2 hours after the beginning numberAmino Diet of the meal. Plasma was obtained from each blood acid123456mg/100 sample by centrifuging at 1,800 X g for 20 minutes at 5°.A portion of the plasma from miValineIsoleucineLancineThreonineMethioninePhenylalanineLysineAlanineGlycine!Proline1SerineAspartic each subject was deproteinized with 1% picric acid, centrifuged, and the picrate removed from the supernatant by ionexchange chromatography; the eluent was analyzed for 16 amino acids by the method of Gehrke et al.7 (36) using a gas chromatograph.8 Transexamic acid (trans-4(aminomethyl)-cyclohexane-carboxylic acid) was used as the internal standard and the acidGlutamic final volatile amino acid derivative pre acidTyrosineHistidine"-'AT'-Methyl-histidine*ArginineTotal pared was the N-trifluoro-acetyl-n-butyl ester. A second portion of plasma was de proteinized with 15% sulfosalicylic acid and centrifuged at 27,000 X g for 15 min utes at 5°.The supernatant was analyzed aminoacidsTotal for lysine, histidine, NT-methylhistidine and indispen A/T-methylhistidine using an ion-exchange aminoacidsTotal sable amino acid analyzer9 according to the dispen method of Speckman et al. (37). Plasma sable aminoacids2.71.42.21.90.41.32.85.62.6«3.0°1.70.88.01.30.7-0.032.137.812.824.02.61.42.12.00.61.23.24.82.6*3.1«2.00.89.31.41.5»0.062.040.713.126.02.81.51.92.20.6 urea nitrogen was determined using the method of Foster and Hohholzer (38). 1These amino acida were determined by the method of Each plasma sample from each subject was Zumwalt et al. (30) using a gas-liquid Chromatograph, except analyzed separately. TABLE 2 Mean fasting plasma amino acid concentrations of men consuming the six experimental diets1•*
abbreviations used to describe these six diets appear in figures 1 and 3. Upon omis sion of histidine or arginine from the amino acid mixtures, diammonium citrate was used to keep the total nitrogen constant. The basal diet was calculated to contain 89 mg of histidine and 175 mg of arginine. Methods. Blood from the antecubital vein of each subject was collected in heparinized tubes before breakfast on the first day of the study, at the end of the adjustment period and on the last day of each 7-day experimental period. Following
RESULTS Fasting plasma amino acid concentra tions. The mean plasma amino acid con centrations of the six subjects before break fast on the seventh day of consuming each of the six experimental diets are presented in table 2. Lysine and histidine concentra tions determined by the ion-exchange or the gas Chromatographie (not shown) methods were approximately the same. The AKmethylhistidine (3-methylhistidine ) con centrations were obtained only from the ion-exchange amino acid analyzer. There fore, the histidine and IVT-methylhistidine values reported are those obtained from the ion-exchange amino acid analyzer, and 7 Gehrke, C. W., Roach, C., Zumwalt, R. W., Stalling, D. L. & Wall, L. L. (1968) Quantitative Gas-Liquid Chromatography of Amino Acids in Proteins and Biological Substances. Analytical Biochemistry Lab oratories, Columbia, Missouri. 8 Bendix model 2500, Bendlx Corporation, Ronceverte, West Virginia. • Beckman model 121, Beckman Instruments, Inc., Splnco Division, Palo Alto, California.
Downloaded from https://academic.oup.com/jn/article-abstract/107/11/2078/4768996 by Tulane University Medical Library user on 31 January 2019
for histidine and #r-methylhistidine (see footnote 5). *Means are for six subjects. BEMvalues for the individual amino acida at O, 1, and 2 hr are presented in figures 2 to 4 and table 3. SEM values for the total amino acids, total indispensable and total dispensable amino acids at 0 hr are 2.4, 0.9, and 1.6, respec tively. » Statistically significant differences at P < 0.05 for glycine and proline and at P < 0.01 for histidine are indicated by small letters; diets sharing a common letter are not sig nificantly different. *Diets 1, 3, and 5 versus diets 2, 4, and 6 are significantly different at P < 0.01. 5 Determined by the method of Spackman et al. (37) using an ion-exchange amino acid analyzer.
2081
HISTIDINE, ARGININE AND PLASMA AMINO ACIDS
1 0.66 2 0.42
b1,3, Diets0 hr:
bs
6«1 4.
vs.05
10 l
2| 1 2A*.b2, 1 21••— 1 1 2A«valuation:». ]2His Hours ifter a mael 4Diet 1 Number, +ArgAStatisticalb0 IAA
RM|
'IAA
+
-0 4-- Ì
25+1
26Hit1
DAC —Arg^»,
Diet ]-HitAa0 Competition + Arg3
+ Arghr
Fig. 1 Mean plasma urea nitrogen levels of six young men fed diets differing in source of nonspecific nitrogen. Abbreviations at the bottom of the figure refer to the mixture of eight indispensable amino acids plus cystine and tyrosine (IAA), of six dispensable amino acids (DAA), glycine (Gly), diammonium citrate (DAC), histidine (His), and arginine (Arg). Means are the average values for all six subjects on the last day of each 7-day experimental period in plasma obtained before breakfast (0 hour) and at 1 and 2 hours following breakfast. Small letters indicating statistical evaluations ( P
