scholarly journals Studies on the biosynthesis in vivo and excretion of 16-unsaturated C19 steroids in the boar

1972 ◽  
Vol 129 (3) ◽  
pp. 657-663 ◽  
Author(s):  
Y. A. Saat ◽  
D. B. Gower ◽  
F. A. Harrison ◽  
R. B. Heap

1. In one experiment [7α-3H]pregnenolone was infused continuously for 12min into the left spermatic artery of a sexually mature boar and blood was collected during this period by continuous drainage from the spermatic vein. After infusion, the testis was removed and immediately cooled to −196°C. 2. From both the testicular tissue and the spermatic venous plasma, 3H-labelled 16-unsaturated C19 steroids were isolated and characterized and their radiochemical purity was established. 5α-Androst-16-en-3α- and 3β-ol occurred mainly as sulphate conjugates and to a lesser extent as free steroids. Only traces of these alcohols occurred as glucosiduronate conjugates. 5α-Androst-16-en-3-one was found in the free (ether-extractable) fraction. 3. The isotope concentration of each of the 3H-labelled 16-unsaturated C19 steroids in testicular tissue was different from that in spermatic venous plasma. 4. The ratios of tritiated 5α-androst-16-en-3α- and 3β-ol (free steroids) to their respective sulphate conjugates in the testicular tissue were less than the ratios of the same compounds in the spermatic venous plasma. The possibility that the sulphates are partially hydrolysed by testicular sulphatases before secretion is discussed. 5. In a second experiment, a continuous close-arterial infusion of [7α-3H]pregnenolone into the left testis was performed over a 200min period and all the urine that accumulated during the infusion was collected for analysis. 6. No 3H-labelled 16-unsaturated C19 steroids were detected in the urine as free steroids. Only a trace of 5α-androst-16-en-3α-ol was detected conjugated as glucosiduronate, whereas the corresponding 3β-alcohol occurred mainly as glucosiduronate and to a lesser extent as sulphate. 7. The absence of 5α-androst-16-en-3β-ol glucosiduronate in the spermatic venous blood and its presence in considerable amount in the urine may be attributed to hepatic glucuronyl transferase activity.

1974 ◽  
Vol 144 (2) ◽  
pp. 347-352 ◽  
Author(s):  
Y. A. Saat ◽  
D. B. Gower ◽  
F. A. Harrison ◽  
R. B. Heap

1. [5α-3H]5α-Androst-16-en-3-one (5α-androstenone) was infused at a constant rate for 180min into the spermatic artery of a sexually mature boar. Samples of spermatic-venous blood were collected at 1min intervals for the first 10min of the infusion and thereafter at 15min intervals for the first hour, then at 64, 125, 155 and 172min. After infusion, the testis was removed and immediately cooled to −196°C. 2. From both the testicular tissue and the spermatic-venous plasma, endogenous and3H-labelled androst-16-enes were isolated, characterized and quantitatively determined and their specific radioactivity was calculated. 3. The specific radioactivities of 5α-androstenore, 5α-androst-16-en-3α-ol and 5α-androst-16-en-3β-ol (an-α and an-β) in testicular tissue were different from those in the spermatic-venous plasma, suggesting that these compounds may be present in more than one compartment of the testis and differentially secreted into the spermatic-venous blood. 4. The ratios of the specific radioactivities of an-α and an-β to their respective sulphate conjugates in the testicular tissue were less than the ratios of the same compounds in the spermatic-venous plasma. 5. The patterns of secretion of these labelled compounds in the spermatic-venous blood during the period of infusion were demonstrated. 6. The urine that accumulated during the infusion was analysed and found to contain3H-labelled an-β, conjugated as both glucuronide and sulphate, the specific radioactivities of which were determined. Little or no androst-16-enes occurred as free steroids. 7. The presence of an-β glucuronide in the urine is discussed.


1984 ◽  
Vol 103 (2) ◽  
pp. 179-186 ◽  
Author(s):  
E. L. Hurden ◽  
D. B. Gower ◽  
F. A. Harrison

ABSTRACT Three mature Large White boars were anaesthetized and received [7(n)-3H]pregnenolone by continuous infusion into right and left spermatic arteries for up to 180 min. Spermatic venous blood flow was measured by separate timed collections of completely diverted outflow from each testis and blood not sampled was returned to the peripheral circulation. The total radioactivity in plasma from each testis increased markedly during the first 60 min of infusion to reach a plateau from 80 to 180 min. Radiolabelling of 5α-androst-16-en-3-one, 5α-androst-16-en-3β-ol and -3α-ol showed similar patterns with ratios of mean radioactivity of 5:3:1 respectively between 80 and 180 min. In comparison, the amounts of tritiated 4,16-androstadien-3-one formed were very small. The radiolabelling of testosterone and 4-androstenedione occurred more rapidly than that of the 16-androstenes and reached maxima by 30 min. However the amounts were only one-fifth (testosterone) and one-tenth (4-androstenedione) those of the combined quantities of tritiated 16-androstenes. Addition of human chorionic gonadotrophin (hCG) to the infusate to one testis in each animal (so that 5000 i.u. hCG were delivered in 15–20 min) produced no change in the outputs of radiolabelled steroids although radioimmunoassay of spermatic venous plasma in samples from the third experiment showed a transient increase in the concentration of 4-androstene-3,17-dione during the hCG infusion. It is suggested the lack of response to hCG could be produced by saturation and down regulation of binding sites by the very high local concentrations of hCG. J. Endocr. (1984) 103, 179–186


1970 ◽  
Vol 46 (1) ◽  
pp. 21-28 ◽  
Author(s):  
M. C. RAHEJA ◽  
O. J. LUCIS

SUMMARY The synthesis of testosterone from [4-14C]dehydroepiandrosterone (DHEA) and [7α-3H]dehydroepiandrosterone sulphate (DHEA-S) by human testes in vivo and in vitro was investigated. Neither free testosterone nor free DHEA was found in a perfused testis or the spermatic venous plasma after the infusion of [7α-3H]DHEA-S into the spermatic artery in vivo, whereas 3H-labelled free DHEA, testosterone and androstenedione were isolated after incubation of testicular tissue with the same substrate in vitro. Only 14C-labelled testosterone was found in the spermatic venous effluent and in the testis after infusion of a mixture of equimolar amounts of [7α-3H]-DHEA-S and [4-14C]DHEA into the spermatic artery in vivo. Testosterone containing 3H and 14C was isolated after incubation of testicular tissue with a mixture of the two substrates in vitro.


1974 ◽  
Vol 61 (3) ◽  
pp. 401-410 ◽  
Author(s):  
H. W. A. de BRUIJN ◽  
H. J. van der MOLEN

SUMMARY 17α,20α-Dihydroxy-4-pregnen-3-one is a competitive inhibitor of C17,20-lyase activity in rat testicular tissue in vitro and the significance of this inhibition in vitro was evaluated for testosterone biosynthesis in rat and rabbit testis in vivo. It is concluded that 17α,20α-dihydroxy-4-pregnen-3-one is not involved in the regulation of C17,20-activity in vivo, because it was not possible to detect any 17α,20α-dihydroxy-4-pregnen-3-one in rat and rabbit testicular tissue or in testicular venous blood. If present, the levels are lower than 10 pmol/g testis. Levels of 17α-hydroxyprogester-one are in the order of 50 pmol/g testis. The C17,20-lyase has a higher affinity for 17α-hydroxyprogesterone than for 17α,20α-dihydroxy-4-pregnen-3-one and hence inhibition under in-vivo conditions is not favoured. In rat testes the 20α-hydroxysteroid dehydrogenase activity, which can convert 17α-hydroxyprogesterone to 17α,20α-dihydroxy-4-pregnen-3-one, was found to be mainly (97%) localized in the seminiferous tubules and not at the site of testosterone formation in the interstitial tissue.


1990 ◽  
Vol 258 (5) ◽  
pp. E888-E893 ◽  
Author(s):  
E. Hagstrom ◽  
P. Arner ◽  
U. Ungerstedt ◽  
J. Bolinder

The in vivo kinetics of lactate and pyruvate in the extracellular space of subcutaneous adipose tissue after glucose ingestion were investigated in healthy volunteers by the use of a microdialysis sampling technique. Comparison was made with the metabolite levels in venous plasma. The absolute subcutaneous tissue concentrations of lactate and pyruvate were estimated in the fasting state by perfusion with varying lactate- and pyruvate-containing solutions. An equilibrium with the surrounding extracellular fluid was found for both lactate and pyruvate in concentrations similar to those in venous plasma. After glucose ingestion there was an increase in the circulating levels of glucose, lactate, and pyruvate, which returned to base-line values within 3 h. There was a more marked increase in lactate in subcutaneous adipose tissue than in venous blood, and the adipose tissue lactate remained elevated for at least 3 h. In contrast, pyruvate levels increased much less in subcutaneous fat than in venous blood. The addition of isoproterenol (which inhibits adipose tissue glucose metabolism) to the tissue perfusate lowered the subcutaneous tissue lactate levels significantly but did not affect the subcutaneous pyruvate levels. These data suggest that human subcutaneous adipose tissue is a source of in vivo lactate production after glucose ingestion. Since lactate is thought to be a major substrate for glycogen synthesis in the liver, the present findings may provide evidence of a new and important role of the adipose tissue metabolism in the regulation of whole body glucose homeostasis in humans.


1991 ◽  
Vol 30 (01) ◽  
pp. 35-39 ◽  
Author(s):  
H. S. Durak ◽  
M. Kitapgi ◽  
B. E. Caner ◽  
R. Senekowitsch ◽  
M. T. Ercan

Vitamin K4 was labelled with 99mTc with an efficiency higher than 97%. The compound was stable up to 24 h at room temperature, and its biodistribution in NMRI mice indicated its in vivo stability. Blood radioactivity levels were high over a wide range. 10% of the injected activity remained in blood after 24 h. Excretion was mostly via kidneys. Only the liver and kidneys concentrated appreciable amounts of radioactivity. Testis/soft tissue ratios were 1.4 and 1.57 at 6 and 24 h, respectively. Testis/blood ratios were lower than 1. In vitro studies with mouse blood indicated that 33.9 ±9.6% of the radioactivity was associated with RBCs; it was washed out almost completely with saline. Protein binding was 28.7 ±6.3% as determined by TCA precipitation. Blood clearance of 99mTc-l<4 in normal subjects showed a slow decrease of radioactivity, reaching a plateau after 16 h at 20% of the injected activity. In scintigraphic images in men the testes could be well visualized. The right/left testis ratio was 1.08 ±0.13. Testis/soft tissue and testis/blood activity ratios were highest at 3 h. These ratios were higher than those obtained with pertechnetate at 20 min post injection.99mTc-l<4 appears to be a promising radiopharmaceutical for the scintigraphic visualization of testes.


1978 ◽  
Vol 87 (1) ◽  
pp. 181-191 ◽  
Author(s):  
Alfred S. Wolf ◽  
Klaus A. Musch ◽  
Werner Speidel ◽  
Jürgen R. Strecker ◽  
Christian Lauritzen

ABSTRACT A new model for the perfusion of human term-placentas has been developed for studies on the placental biogenesis of C-18 and C-19 steroids. For viability criteria, the glucose- and oxygen-consumption, regional perfusion control by dye-infusions or scanning after injection of 99Tc-labelled macroparticles, and the histological qualification were chosen. The recycled perfusate was investigated for the steroids oestrone (Oe1), oestradiol-17β (Oe2), oestriol (Oe3), 4-androstene-3,17-dione (A), testosterone (T), and human placental lactogen (HPL) by radioimmunoassay in controls and perfusions with the foetal steroid precursor dehydroepiandrosterone sulphate (DHA-S). In control perfusions, steroid hormones were found in constant ratios (Oe1:Oe2:Oe3:T:A = 30:1.5:100:0.35:1). Following the administration of 10 mg DHA-S for testing the metabolic capacity of the organ, high concentrations of Oe1 (90–720 ng/ml = 250–3970 % as compared to 100% pre-injection values) were found, shortly preceded by a rapid increase of A (66–1000 ng/ml = 100–16 000 %). A typical surge of T (5.3–147 ng/ml = 265–4640 %) preceded the normally slower increment of Oe2 (22–220 ng/ml = 1570–4330 %). The concentrations of Oe3 and HPL remained nearly unchanged. From different steroid patterns after DHA-S-load, two distinct responses of term-placentas could be differentiated: Group I (n=12) showed high concentrations of Oe1 (3200 ± 940 %), a small increase of T (1020 ± 500%), as well as low and delayed values of Oe2 (1660 ± 450%). In Group II (n = 5), values were high for T (3160 ± 1020%) and Oe2 (3300 ± 1110%), whereas Oe1 was found in a lower range (508 ± 302%). In contrast to in vivo findings in maternal venous blood after DHS-S injection to the mother, oestrone was found in perfusions as the main oestrogen fraction from DHA-S. Thus, the analysis of such metabolic differences might be of help in the interpretation of complex results from the DHA-S-loading test.


Sensors ◽  
2021 ◽  
Vol 21 (11) ◽  
pp. 3827
Author(s):  
Gemma Urbanos ◽  
Alberto Martín ◽  
Guillermo Vázquez ◽  
Marta Villanueva ◽  
Manuel Villa ◽  
...  

Hyperspectral imaging techniques (HSI) do not require contact with patients and are non-ionizing as well as non-invasive. As a consequence, they have been extensively applied in the medical field. HSI is being combined with machine learning (ML) processes to obtain models to assist in diagnosis. In particular, the combination of these techniques has proven to be a reliable aid in the differentiation of healthy and tumor tissue during brain tumor surgery. ML algorithms such as support vector machine (SVM), random forest (RF) and convolutional neural networks (CNN) are used to make predictions and provide in-vivo visualizations that may assist neurosurgeons in being more precise, hence reducing damages to healthy tissue. In this work, thirteen in-vivo hyperspectral images from twelve different patients with high-grade gliomas (grade III and IV) have been selected to train SVM, RF and CNN classifiers. Five different classes have been defined during the experiments: healthy tissue, tumor, venous blood vessel, arterial blood vessel and dura mater. Overall accuracy (OACC) results vary from 60% to 95% depending on the training conditions. Finally, as far as the contribution of each band to the OACC is concerned, the results obtained in this work are 3.81 times greater than those reported in the literature.


2006 ◽  
Vol 95 (03) ◽  
pp. 434-440 ◽  
Author(s):  
Satu Hyytiäinen ◽  
Ulla Wartiovaara-Kautto ◽  
Veli-Matti Ulander ◽  
Risto Kaaja ◽  
Markku Heikinheimo ◽  
...  

SummaryThrombin regulation in newborns remains incompletely understood.We studied tissue factor-initiated thrombin formation in cord plasma in vitro, and the effects of Factor VLeiden (FVL) heterozygosity on thrombin regulation both in vitro and in vivo in newborns. Pregnant women with known thrombophilia (n=27) were enrolled in the study. Cord blood and venous blood at the age of 14 days were collected from 11 FVL heterozygous newborns (FVL-positive) and from 16 FVL-negative newborns. Prothrombin fragment F1+2 and coagulation factors were measured. Tissue factor-initiated thrombin formation was studied in cord platelet-poor plasma (PPP) of FVL-negative and -positive newborns, and in both PPP and platelet-rich plasma (PRP) of healthy controls. The endogenous thrombin potential (ETP) in cord PPP or PRP was ∼60% of that in adult plasma, while thrombin formation started ∼55% and ∼40% earlier in cord PPP and PRP, respectively. Further, in FVL-positive newborns thrombin formation started significantly earlier than in FVL-negative newborns. Exogenous activated protein C (APC) decreased ETP significantly more in cord than in adult PRP. In FVL-negative cord plasma 5nM APC decreased ETP by 17.4±3.5% (mean±SEM) compared with only 3.5±3.8% in FVL-positive cord plasma (p=0.01). FVL-positive newborns showed similar levels of F1+2 but significantly decreased levels of factor V compared with FVL negative newborns both in cord plasma (FV 0.82±0.07 U/ml vs. 0.98±0.05 U/ml, p=0.03) and at the age of two weeks (FV 1.15±0.04 U/ml vs. 1.32±0.05 U/ml, p=0.03). In conclusion, newborn plasma showed more rapid thrombin formation and enhanced sensitivity to APC compared with adult plasma. FVL conveyed APC resistance and a procoagulant effect in newborn plasma. Lack of elevated F1+2 levels in FVL-positive infants, however, suggested the existence of balancing mechanisms; one could be the observed lower level of factor V in FVL heterozygous newborns.


1978 ◽  
Vol 79 (2) ◽  
pp. 179-190 ◽  
Author(s):  
MRINAL K. SANYAL

The concentrations of progesterone and 5α-pregnane-3,20-dione in ovarian and uterine venous plasma and in the systemic circulation were measured during gestation in the rat. The steroids were quantified by radioimmunoassay after separation on silicic acid microcolumns with the solvent system hexane: ethyl acetate (5: 2, v/v). The concentration of progesterone in the systemic circulation was highest on days 3–4 and 13–17 of pregnancy; throughout gestation, the concentration of 5α-pregnane-3,20-dione was low in relation to that of progesterone and showed no marked changes as gestation proceeded. The level of progesterone in ovarian venous effluent was 10–20 times higher than that in the uterine vein and 20–50 times greater than that in the systemic circulation. The rate of secretion of progesterone by the ovary was highest during days 13–17 of gestation and ovariectomy during this period markedly reduced the levels of progesterone in the peripheral circulation. The concentration of progesterone in the uterine venous effluent was raised compared with the concentration in plasma from the abdominal aorta, especially on days 7 and 9 of pregnancy. These results suggest that, in vivo, the rat placenta synthesizes small amounts of progesterone and secretes it into the maternal circulation. The ovary is the primary source of progesterone during pregnancy and the placental contribution is of secondary importance. Although 4-ene-5α-reductase enzyme(s) is present in the ovary and placenta, significant quantities of the reduced progestin 5α-pregnane-3,20-dione are not secreted into the systemic circulation during gestation in the rat.


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