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Urinary testosterone in high altitude natives
351
URINARY TESTOSTERONE IN HIGH ALTITUDE NATIVES
Roger Guerra-Garcia, Alicia Velasquez and Jo&
Whittembury
Instituto de Investigaciones de la Altura Universidad Peruana Cayetano Heredia Lima, PerG Received June 21, 1965 ABSTRACT Urinary testosterone was measured using thin layer and gas-liquid chromatography in a group of eight men between 21-25 years of age, born and living at 14,300 ft. The mean 24 hours excretion was 96.5 pg k 10.16 (S.E.), similar to the mean of ten men of the same age range, natives from and living at sea level: 99.8 f 14.3 (S.E.). There was no correlation between body surface and the excretion of testosterone in these groups. It has
been described that acute exposure to high
altitude produces reversible damage to the seminipherous epithelium of sheep and guinea pigs with a return to normal ten weeks later (1,2,3). modification.
The intersitial cells did not show any
To observe whether similar changes occur in
man upon acute exposure to high altitude,a group of volunteers has been studied in regard to various endocrine functions related to reproduction. This paper reports the findings in urinary testosterone of natives of high altitude and the control values of the men
from sea level.
SUBJECTS Eight normal men, of ages betueen 21-25 years, born and
352
STEROIDS
6:3
living in Cerro de Pasco (altitude 14,300 ft.) were studied. The mean barometric pressure was 450 mm Hg in this place. Physical examination showed no abnormality of the sexual area and the spermatogram was normal. As controls we had ten volunteers from the Peruvian Navy, men of ages between 19-22 years, natives from and living at sea level. METHODS We have used the procedure of Ibayashi -et al (4), in brief: to one fourth of a 24-hours urine collection 0.02 kc of 1.2,3H-testosterone (s.a. 135 cLc/Kg) was added as internal standard, then enzymatic hydrolisis, ether extraction and thin layer chromatography in benzene-ethyl acetate (1:l) were performed. A testosterone standard and the eluate from the first TLC were acetylated with acetic anhydride 0.1 ml and pyridine 0.1 ml at roam temperature, for 18 hours. The excess reagents were evaporated under nitrogen: preliminary work demonstrates that the reaction is quantitative. The acetylated extract was developed in TLC using a benzene-ethyl acetate (3:l) system. The running time was 75 minutes for each chromatogram (solvent front moved 15 cm). Silica gel was CAMAG, grade D5F. Gas Chromatosraohv A Research Specialties Co., Model 600 with an Argon Detector was used. Column: 6 ft x l/16 inch i.d. stainless steel. Phase: 3% SE-30 on Diatoport "S" 80-100 mesh. Carrier gas: Argon. Flowrate at room temperature: 60 ml/min at 25 lbs/sq inch. Detector temperature: 25OoC. Column temperature: 2450. Vaporizer temperature: 27OoC. The eluate from the second TLC was dissolved in 50 ~1 of acetone, and 5 ~1 of this solution were injected with a Hamilton syringe into the column. The remaining solution was used to check the recovery of the added 3H-testosterone using the 6000 Dynacon Electrometer system of the Nuclear Chicago Corp. Quantification of the peak was carried out by comparing the testosterone acetate peak height of the sample with that of a standard. A linear response of the detector in the range of concentration selected (0.5-2.5 pg) was also demonstrated. RESULTS AND DISCUSSION Table I summarizes the results of these experiments,
73,2 68,O 97,Y
1,62
1,48
1,70
1,60
23
21
22
23
N.A.
A.A.
W.P.
Mean
+
S.P.
79,s
1,54
25
E.A.
96,5
O.R.
113,l
1,60
22
V.B.
71,O 99,8 + 14.3
1,48 1,62
20
F.F.
20
17790
I,63
22 G.C.
10.1 (S.E.)Mean
11396
5392
17690
10796
1,56
1951
1,61
1,76
80,O
20
20
19
21
1,69
H.M.
R.V.
V.R.
21
7290
1,62
19
R.M.
10436
1,65
24
A.O.
9690
1,65
19
15595
1,67
25
D.C.
V.C.
5096
1,59
5290
TESTOSTERONE ug / 24 hr
1,66
BODY SURFACE sq meters
19
AGE
SEA LEVEL SUBJECTS
A.V.
SUBJECTS
25
BODY SURFACE TESTOSTERONE sq meters w / 24 hr
V.A.
SUBJECTS AGE
HIGH ALTITUDE SUBJECTS
URINARY TESTOSTERONE
TABLE I
(S.E.) ifI
w
STEROIDS
354
6:3
they show no difference between the groups. An indication that efficient adaptation to an altered environment has been achieved, is the ability to multiply normally, and for this purpose, testosterone secretion plays an important role.
These results suggest that the testis
of high altitude natives has an endocrine function similar to that of sea level subjects.
Our results agree well with
the normal range of testosterone reported from other laboratories (4,5,6) but to our knowledge it is the first series reported in a homogenous group with respect to age.
The
range of urinary testosterone excretion of subjects at high altitude, 51-155 Kg/24 hr, was almost as wide as the normal range of Ibayashi's
(4) series; 41-200 pg/24 hr, although age range
in his series was from 22-42 years. We tried to find a direct correlation between the excretion of testosterone and the body surface area but this could not be domonstrated statistically.
(r = + 0.43 + 0.34
S-E.). ACKNOWLEDGEMENTS The authors are grateful to Dr. Herbert H. Wotiz for the loan of part of the equipment used in this work. Miss Gladys Silva kindly helped us with the manuscript. Supported by Grants TWO0096 and 08576 from the National Institute of Health, United States Public Health Service. REFERENCES 1.
M., Atkins, J. and Castason, J., Monge, C., San Mar&, AN. FAC. MED. LIMA 28, 15 (1945).
Sept. 1965
STEROIDS
355
2.
Altland, P.D., PHYSIOL. ZOOL. 22, 235 (1949).
3.
Guerra-Garcza, R., Bachellor's Thesis, San Marcos University School of Medicine, Lima (1959).
4.
Ibayashi, H., Nakamura, M., Murakawa, S., Uchikawa, T., Tanioka, T. and Nakao, K., STEROIDS 1, 559 (1964).
5.
Camacho, A.M. and Migeon, C.J., J. CLIN. ENDOCRINOL. 23, 301 (1963).
6.
Futterweit, W., McNiven, N-L., Guerra-Garcia, R., Gibree, N., Drosdowsky, M., Siegel, G-L., Soffer, L.J., Rosenthal, I.M. and Dorfman, R.I., STEROIDS 4, 137 (1964).
URINARY TESTOSTERONE IN HIGH ALTITUDE NATIVES
Roger Guerra-Garcia, Alicia Velasquez and Jo&
Whittembury
Instituto de Investigaciones de la Altura Universidad Peruana Cayetano Heredia Lima, PerG Received June 21, 1965 ABSTRACT Urinary testosterone was measured using thin layer and gas-liquid chromatography in a group of eight men between 21-25 years of age, born and living at 14,300 ft. The mean 24 hours excretion was 96.5 pg k 10.16 (S.E.), similar to the mean of ten men of the same age range, natives from and living at sea level: 99.8 f 14.3 (S.E.). There was no correlation between body surface and the excretion of testosterone in these groups. It has
been described that acute exposure to high
altitude produces reversible damage to the seminipherous epithelium of sheep and guinea pigs with a return to normal ten weeks later (1,2,3). modification.
The intersitial cells did not show any
To observe whether similar changes occur in
man upon acute exposure to high altitude,a group of volunteers has been studied in regard to various endocrine functions related to reproduction. This paper reports the findings in urinary testosterone of natives of high altitude and the control values of the men
from sea level.
SUBJECTS Eight normal men, of ages betueen 21-25 years, born and
352
STEROIDS
6:3
living in Cerro de Pasco (altitude 14,300 ft.) were studied. The mean barometric pressure was 450 mm Hg in this place. Physical examination showed no abnormality of the sexual area and the spermatogram was normal. As controls we had ten volunteers from the Peruvian Navy, men of ages between 19-22 years, natives from and living at sea level. METHODS We have used the procedure of Ibayashi -et al (4), in brief: to one fourth of a 24-hours urine collection 0.02 kc of 1.2,3H-testosterone (s.a. 135 cLc/Kg) was added as internal standard, then enzymatic hydrolisis, ether extraction and thin layer chromatography in benzene-ethyl acetate (1:l) were performed. A testosterone standard and the eluate from the first TLC were acetylated with acetic anhydride 0.1 ml and pyridine 0.1 ml at roam temperature, for 18 hours. The excess reagents were evaporated under nitrogen: preliminary work demonstrates that the reaction is quantitative. The acetylated extract was developed in TLC using a benzene-ethyl acetate (3:l) system. The running time was 75 minutes for each chromatogram (solvent front moved 15 cm). Silica gel was CAMAG, grade D5F. Gas Chromatosraohv A Research Specialties Co., Model 600 with an Argon Detector was used. Column: 6 ft x l/16 inch i.d. stainless steel. Phase: 3% SE-30 on Diatoport "S" 80-100 mesh. Carrier gas: Argon. Flowrate at room temperature: 60 ml/min at 25 lbs/sq inch. Detector temperature: 25OoC. Column temperature: 2450. Vaporizer temperature: 27OoC. The eluate from the second TLC was dissolved in 50 ~1 of acetone, and 5 ~1 of this solution were injected with a Hamilton syringe into the column. The remaining solution was used to check the recovery of the added 3H-testosterone using the 6000 Dynacon Electrometer system of the Nuclear Chicago Corp. Quantification of the peak was carried out by comparing the testosterone acetate peak height of the sample with that of a standard. A linear response of the detector in the range of concentration selected (0.5-2.5 pg) was also demonstrated. RESULTS AND DISCUSSION Table I summarizes the results of these experiments,
73,2 68,O 97,Y
1,62
1,48
1,70
1,60
23
21
22
23
N.A.
A.A.
W.P.
Mean
+
S.P.
79,s
1,54
25
E.A.
96,5
O.R.
113,l
1,60
22
V.B.
71,O 99,8 + 14.3
1,48 1,62
20
F.F.
20
17790
I,63
22 G.C.
10.1 (S.E.)Mean
11396
5392
17690
10796
1,56
1951
1,61
1,76
80,O
20
20
19
21
1,69
H.M.
R.V.
V.R.
21
7290
1,62
19
R.M.
10436
1,65
24
A.O.
9690
1,65
19
15595
1,67
25
D.C.
V.C.
5096
1,59
5290
TESTOSTERONE ug / 24 hr
1,66
BODY SURFACE sq meters
19
AGE
SEA LEVEL SUBJECTS
A.V.
SUBJECTS
25
BODY SURFACE TESTOSTERONE sq meters w / 24 hr
V.A.
SUBJECTS AGE
HIGH ALTITUDE SUBJECTS
URINARY TESTOSTERONE
TABLE I
(S.E.) ifI
w
STEROIDS
354
6:3
they show no difference between the groups. An indication that efficient adaptation to an altered environment has been achieved, is the ability to multiply normally, and for this purpose, testosterone secretion plays an important role.
These results suggest that the testis
of high altitude natives has an endocrine function similar to that of sea level subjects.
Our results agree well with
the normal range of testosterone reported from other laboratories (4,5,6) but to our knowledge it is the first series reported in a homogenous group with respect to age.
The
range of urinary testosterone excretion of subjects at high altitude, 51-155 Kg/24 hr, was almost as wide as the normal range of Ibayashi's
(4) series; 41-200 pg/24 hr, although age range
in his series was from 22-42 years. We tried to find a direct correlation between the excretion of testosterone and the body surface area but this could not be domonstrated statistically.
(r = + 0.43 + 0.34
S-E.). ACKNOWLEDGEMENTS The authors are grateful to Dr. Herbert H. Wotiz for the loan of part of the equipment used in this work. Miss Gladys Silva kindly helped us with the manuscript. Supported by Grants TWO0096 and 08576 from the National Institute of Health, United States Public Health Service. REFERENCES 1.
M., Atkins, J. and Castason, J., Monge, C., San Mar&, AN. FAC. MED. LIMA 28, 15 (1945).
Sept. 1965
STEROIDS
355
2.
Altland, P.D., PHYSIOL. ZOOL. 22, 235 (1949).
3.
Guerra-Garcza, R., Bachellor's Thesis, San Marcos University School of Medicine, Lima (1959).
4.
Ibayashi, H., Nakamura, M., Murakawa, S., Uchikawa, T., Tanioka, T. and Nakao, K., STEROIDS 1, 559 (1964).
5.
Camacho, A.M. and Migeon, C.J., J. CLIN. ENDOCRINOL. 23, 301 (1963).
6.
Futterweit, W., McNiven, N-L., Guerra-Garcia, R., Gibree, N., Drosdowsky, M., Siegel, G-L., Soffer, L.J., Rosenthal, I.M. and Dorfman, R.I., STEROIDS 4, 137 (1964).