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Biomedical subjects

H Degrelle

Publications and source records attributed to H Degrelle.

At least 37 records · Page 2Linked to original sources

Opponent strain effect on eliciting attacks in NZB mice: physiological correlates.

In agonistic encounters between male mice, the characteristics of the opponent may influence the attacking behavior of its partner. The present study shows that the opponent's ability to elicit attacking behavior in NZB males is strain dependent. BALB/c opponents elicit attacks more frequently, earlier and more intensively than C57BL/6 males. Plasma testosterone concentration was found to be higher in BALB/c than in C57BL/6 intact males. The weight of seminal vesicles in castrated males of both strains increased with injections of either 10- or 250-micrograms testosterone propionate (TP). This response was greater in BALB/c with the higher TP dose. The submandibular glands reacted to TP only in castrated BALB/c males with the higher dose. Furthermore, BALB/c males produced more marking secretions than C57BL/6 males. These results suggest that for these two strains, a higher testosterone sensitivity and a greater production of secretions are associated with a higher probability of opponents to elicit attacks. Genetic hypotheses on the underlying mechanisms are discussed.

Aggression↗

Internalization of human sex steroid-binding protein in the monkey epididymis.

Human sex steroid-binding protein (hSBP) has been purified from late-pregnancy serum and labelled either by iodination (125I) or by photoaffinity with [3H]delta 6-testosterone. Using a micromanipulator, each labelled protein was separately injected into the lumen of epididymal tubules isolated from the head epididymis of the cynomologus monkey (Macaca fascicularis). Tubules were sampled from 3 to 90 min after the injection and processed for electron microscope autoradiography. Localization of the label occurred over the epididymal epithelium whichever tracer was used. The labelling was not randomly distributed over the different cell types constituting the epithelium, since only the 'principal cells' exhibited a silver grain count significantly greater than the background count. In these cells, labelled protein was found over endocytic organelles (coated structures, endosomes, multivesicular bodies and the trans Golgi network) and nuclei (including the nuclear envelope). Quantitative analysis demonstrated the same pattern of cellular and subcellular distribution for each tracer. Pretreatment with excess unlabelled protein significantly reduced the uptake of radioactivity by the principal cells, demonstrating the specificity of this phenomenon. This is the first study to show direct histological evidence for the internalization of hSBP in the primate epididymis, consistent with earlier immunohistochemical or biochemical localization of this protein. It is concluded that head epididymal cells are able to take up labelled hSBP across their apical membrane. The mechanism of internalization seems to involve endocytosis by the principal cells and leads to labelling of the nuclear compartment. This is strikingly similar to the pattern of uptake of rat androgen-binding protein (rABP) by rat epididymal cells previously demonstrated by our group. To what extent the chemical and structural homology between hSBP and rABP can be held responsible for the common cytophysiological behaviour of these sex steroid-binding proteins remains to be determined.

Affinity Labels↗

Effect of testosterone on regulation of the level of sex steroid-binding protein mRNA in monkey (Macaca fascicularis) liver.

Two specific oligonucleotide probes complementary to different regions of human sex steroid-binding protein (SBP) cDNA were used to study the levels of hepatic monkey SBP mRNA during different hormonal states. In females the SBP mRNA level was higher than in males and paralleled the serum SBP level. After castration, the SBP concentration increased in the serum but was reduced after testosterone treatment. In contrast, the hepatic SBP mRNA level decreased after castration and was restored by testosterone treatment. These results suggest a high homology of the nucleotide sequence between human and monkey SBP mRNAs. The changes in liver SBP mRNA levels may explain the sex difference in plasma SBP concentrations, but mechanisms other than the regulation of transcription may regulate the plasma concentration in monkeys.

Animals↗

Immunochemical characterization of corticosteroid-binding globulin in human bronchoalveolar fluid.

The corticosteroid-binding globulin (CBG) is a plasma protein which is present both in liver, where it is mainly synthesized, and in cells of different target tissues for glucocorticoids. Using monospecific antibodies raised against human CGB, we could demonstrate the antigenic identity of the protein in human bronchoalveolar fluid. We found that the bronchoalveolar fluid/serum concentration ratio of CBG was similar to that of albumin. Since albumin is not synthesized in pulmonary cells, it was concluded that, in healthy human, CBG enters bronchoalveolar fluid by diffusion through alveolar cells. It is suggested that the expression of the CBG gene in pulmonary cells could occur during the pathological state.

Blood Proteins↗

Sex steroid-binding protein: identification and comparison of the primary product following cell-free translation of human and monkey (Macaca fascicularis) liver RNA.

A very close similarity in molecular, steroid-binding and immunological properties have been demonstrated for the sex steroid-binding proteins of plasma from human (hSBP) and monkey (mSBP): both are glycoproteins composed of two similar subunits able to bind one steroid molecule and to cross-react with the same antibodies. After translation of human and monkey (Macaca fascicularis) liver mRNAs by a wheat-germ embryo extract, in the presence of labelled amino-acids, we have characterized in both cases a single radioactive polypeptide immunologically related to SBP, migrating in SDS-PAGE as a single band and having a molecular weight of about 42,000. This protein could be displaced from the antibody by pure unlabelled SBP in excess. The difference in molecular weight between the in vitro translation product and the native SBP sub-unit is probably due to the absence of glycosylation in the neo-synthesized protein. The radioactivity incorporated into mSBP was 4 times higher than the radioactivity incorporated into hSBP, suggesting that the amount of mRNA for SBP is higher in monkey than in human liver. Our results show that the two sub-units of hSBP and mSBP derive from a common precursor, representing respectively 0.0050% and 0.0013% of the total neosynthesized proteins in monkey and in human liver.

Animals↗

Effect of the Tfm mutation on handedness in mice.

The hyposthesis has been proposed that testosterone is involved in the determination of handedness in man: a high sensitivity to testosterone being associated with left handedness. Handedness in mice is tested according to Collins' paradigm: most mice present either a right or a left paw preference but others are ambilateral. The hypothesis that there is an association between a low neonatal imprinting by testosterone and a strong handedness (right or left) is tested here using Tfm male mice which are testosterone insensitive. Our results confirmed the hypothesis, since Tfm males were as well lateralized as their female siblings and significantly more strongly lateralized than their male siblings not carrying the mutation.

Androgen-Insensitivity Syndrome↗

Ultradian, circadian and seasonal variations of plasma progesterone and LH concentrations during the luteal phase.

The circadian variations in plasma progesterone (P) and LH concentrations were investigated in six women, aged 23-40 years. All were studied in the mid-luteal phase (7 +/- 2 days after LH mid-cycle surge). Experiments were conducted in autumn and in spring. Blood samples were obtained every 15 min for 24 hr. Plasma P and LH concentrations were measured by RIA. Each subject's time-series was analysed using three methods; visual inspection (chronogram), spectral analysis to estimate component periods of rhythms (tau) and cosinor analysis to quantify the rhythms parameters. Marked temporal variations in plasma P concentration were observed in each subject. The maximal variations over a 24-hr period, ranged between 13-58.5 mmol/l. Differences related to sampling time were statistically validated by ANOVA (p less than 0.00001). Significant harmonic periods were detected by spectral analysis but differed among subjects. In all subjects but one, a circadian rhythm was detected. The acrophase location was similar (about 0700 hr) in the four subjects studied in autumn, but ranged from 1940 to 0320 hr in those studied in spring. An ultradian rhythm with tau = 8 hr was also validated in six time-series with similar acrophases (about 0200, 1000, and 1800 hr). Cosinor analysis of pooled data revealed that the 24-hr, 12-hr, and 8-hr rhythms were statistically significant (p = 0.001) in autumn. algebraic sum of these three cosine functions yielded a circadian waveform with peak-times occurring near 0300 and 1130 hr and a trough-time about 2200 hr. In spring, the circadian pattern appeared quite different, and peak-times were found near 0700 and 2000 hr, and trough-times near 0300 and 1500 hr. Furthermore, the 24-hr mean of P was higher in autumn (28.9 +/- 0.4 nmol/l) than in spring (17.2 +/- 0.4 nmol/l), p from ANOVA less than 0.00001. The evidence for a similar circadian LH pattern is not as strong. Seasonal, circadian and ultradian rhythms characterize the physiologic time structure of plasma P concentration in mid-luteal phase.

Activity Cycles↗

[Secretion and transport of androgens in women].

Androgens have a metabolism particularly complex in women on account of their multiple origins. The author points out the transport proteins whose biological roles are not completely known and ascertains that the free hormone concept has not at the present time a biological signification well defined.

Androgens↗

Identification of the primary translation product of the sex steroid-binding protein from monkey liver mRNA in a cell-free system.

The synthesis of monkey (Macaca fascicularis) Sex steroid-Binding Protein (mSBP) in a wheat germ cell-free system in response to liver RNA was demonstrated by use of a specific antiserum raised against purified native human SBP. Antibodies precipitate a single translation product behaving as a 42 kDa protein in sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Western blots of monkey sera subjected to SDS-PAGE and immunorevelation show that the native mSBP migrates as 2 molecular species (50 and 53 kDa) present in the approximate ratio of 1:10, respectively. The difference in apparent molecular weights of the primary translation product and the reduced mature mSBP may represent glycosylation that occurs post translationally. We describe for the first time the biosynthesis of mSBP at the molecular level and suggest that both components of mSBP derive from a common differentially processed precursor. Its mRNA is poorly represented, since the neosynthesized mSBP represents about 0.005% of the total proteins encoded by liver mRNA.

Animals↗

Regulation of plasma sex-steroid-binding protein in adult cynomolgus monkeys (Macaca fascicularis) during different reproductive states.

Levels of the sex-steroid binding protein (mSBP) have been characterized in Cynomolgus monkeys, during different stages of reproduction and under hormonal treatments, by an immunoassay allowing a specific and accurate measurement of the protein itself. Using an antiserum specific for native human SBP, we have determined the mSBP level by electroimmunodiffusion. This method correlates closely with the binding capacity measured by a steady-state polyacrylamide electrophoresis. The levels are lower in males (M = 88 +/- 8.6 nmoles/1, n = 6) than in females in the follicular phase (M = 123 +/- 4.4 nmoles/1, n = 5, p less than 0.001) and in castrated males (M = 172 +/- 13.3 nmoles/1, n = 3, p less than 0.001). During gestation, the SBP level decreases (M = 69 +/- 16.3 nmoles/1, n = 7, in the latter part of pregnancy), but during lactation, it is similar to follicular values. Estrogen treatment fails to increase SBP levels in castrated animals, but the values are reduced by testosterone treatment. Since these results are different from those observed in women, we question the validity of using monkeys as models for understanding the mechanisms controlling the concentration of SBP in human blood.

Animals↗

Immunochemical characterization and quantification of the sex steroid-binding protein (SBP) in human amniotic fluid.

The sex steroid-binding protein (SBP) is a plasma protein whose concentration in the maternal circulation increases during pregnancy. Using monospecific antibodies raised against human SBP, we could demonstrate the antigenic identity of the protein in human amniotic fluid. In this fluid, we found that the SBP concentration was correlated with the total protein concentration throughout gestation. The concentration gradient of SBP between maternal serum and amniotic fluid was compared to that of other serum proteins, in relation to their relative molecular mass, and it was concluded that SBP enters amniotic fluid in a non-specific manner similar to that of other serum proteins. It is suggested that SBP could act to sequester the sex steroid hormones in amniotic fluid.

Amniotic Fluid↗

Pregnenolone metabolism in Y-1 mouse adrenal cells: HPLC analysis and identification of metabolites by gas chromatography-mass spectrometry.

The Y-1 mouse adrenocortical tumor cells secrete into the culture medium some metabolites of pregnenolone. The long term basic steroid secretion was studied for 24 h by high performance liquid chromatography (HPLC). After 6 h incubation, only one chromatographic peak was shown to be quantitatively important. When the cells were incubated with tritium labeled pregnenolone and carrier, the analysis of radioactivity of each HPLC fraction indicated a rapid incorporation of the exogenous precursor and successive secretion/reabsorption kinetics of the metabolites. The final product, accumulated in the medium, was characterized by gas chromatography-mass spectrometry as 11 beta,20 alpha-dihydroxy-4-pregnen-3-one.

Adrenal Glands↗

[Assay of chorionic gonadotrophin, follicle-stimulating hormone and alpha-fetoprotein in the efferent blood in germinal tumors of the testis].

Assays of human chorionic gonadotrophin (hCG), follicle-stimulating hormone (FSH) and alpha-foetoprotein were performed in peripheral blood and spermatic vein in 73 cases of testicular germ cell tumors. This study showed that the detection sensitivity of hCG is up to 40 times higher in the efferent blood of the tumor than in peripheral blood and that the drop in FSH concentrations often precedes a trophoblastic transformation. These results are of particular interest in the search for a minor trophoblastic or embryonic component. As a result, we propose a new strategy leading to a more efficient diagnosis and an adequate therapy.

Adult↗

[De-etherification of estradiol 3-propyl ether by subcellular fractions of rat liver].

The incubation of PE2 ((6,7-3H)-labelled 3-propyl ether of estra-1,3,5(10)triene-3, 17 beta-diol (estradiol)) with various subcellular fractions of rat liver indicated that the hepatic metabolism of this compound occurs mainly in the microsomal fraction. In addition to the formation of 3-propyl ethers of estra-1,3,5(10)triene-3-ol-17-one (estrone) and estra-1,3,5(10-triene-3, 16alpha, 17 beta-triol (estriol) directly deriving from PE2, the microsomal proteins carried out the deetherification of the propyl ether group leading to phenolic steroids; among them, estradiol, estrone, and estriol were characterized. Protein-bound and water-soluble metabolites were found; the effects of glutathione and of the incubation conditions were in agreement with the thioconjugation of these derivatives. The microsomal metabolism of PE2, and specially the deetherification reaction, required the presence of oxygen and of NADPH as cofactor, the optimum pH ranging from 7.4 to 8. The participation of cytochrome P450 in these metabolic pathways was shown by a partially inhibited catabolism with carbon monoxide and by a more active metabolism in males than in females and when animals were pretreated with phenobarbital. These results allowed us to conclude that the hepatic deetherification of PE2 is carried out by a microsomal oxidative system which is very similar to the system involved in the demethylation of methyl ethers of estrogens.

Animals↗

Immunochemical characterization and quantitation of human sex steroid binding plasma protein.

The interest in the measurement of human sex steroid binding plasma protein (h-SBP) is now increasing since it allows the estimation of the free fraction of circulating hormones in plasma. Up to this date, this protein could only be determined by measuring the total binding capacity of serum for dihydrotestosterone (DHT). The purpose of the present work was to purify the protein, to prepare a rabbit monospecific antiserum and to develop an immunoelectrophoretic assay of h-SBP. The immunological assay is specific, accurate and sensitive. A good correlation with the radioligand assay was found. The h-SBP levels obtained by immunoelectrophoretic assay of different serum samples were 5.3 +/- 1.4 (SEM) mg/L in normal men and 13.4 +/- 2.6 (SEM) mg/L in normal women.

Animals↗