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C Benassayag

Publications and source records attributed to C Benassayag.

At least 19 recordsLinked to original sources
PubMed2000

Evidence for progesterone receptors in the human fetoplacental vascular tree.

The presence of progesterone receptors (PR) throughout the human term fetoplacental vascular tree was investigated. By reverse transcription-polymerase chain reaction (RT-PCR), we showed expression of PR mRNAs in stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins. Binding studies and Scatchard analysis revealed a single class of high-affinity binding sites for (3)H-R5020 (promegestone) in cytosolic extracts of all placental vessels, with K(d) values in the range of 2.5-4 nM. High levels of PR were detected in placental vessels compared to other vascular tissues. Thus, maximum binding capacities of stem villi vessels, chorionic arteries and veins, and umbilical arteries and veins were 247 +/- 25, 377 +/- 58, 295 +/- 40, 371 +/- 118, and 672 +/- 144 fmol/mg protein, respectively. Endothelial cell elimination in chorionic arteries did not significantly modify the number of PR. RT-PCR and binding studies also assessed PR expression in cultured placental vascular smooth muscle cells isolated from stem villi vessels. All these data suggested that most of the PR of fetoplacental vessels were from the media. In conclusion, we report here the first evidence of the presence of PR in the muscular layer of human term fetoplacental vessels. This finding, together with the high progesterone concentrations in cord blood, suggests that the interactions between the PR and its ligand may play a role in the physiology and physiopathology of human fetoplacental vascularization.

Adult↗
PubMed1999

Estrogen receptors (ERalpha/ERbeta) in normal and pathological growth of the human myometrium: pregnancy and leiomyoma.

The distributions of the mRNAs for estrogen receptors (ERalpha and ERbeta) and their binding properties in myometria of pregnant and nonpregnant women and in leiomyoma were studied. RT-PCR analysis indicated that the term pregnancy myometria had little ERalpha mRNA, whereas the amounts of ERbeta mRNAs in pregnant or nonpregnant myometria appeared to be similar. Both ERalpha and ERbeta mRNA were greater in certain leiomyoma than in normal nonpregnant myometria. The binding kinetics revealed that two specific binding sites (with high or low affinity) for 17beta-estradiol were present in the nonpregnant myometrium. Only the low-affinity binding sites were detectable in late-pregnancy myometria and in leiomyoma, and their capacities were increased two- to threefold (P < 0.001) in leiomyoma. The pregnancy- and leiomyoma-related changes in myometrial ER status, especially the low concentration of ERalpha mRNA and the lack of high-affinity ER in pregnant women, plus the increased ERalpha and ERbeta mRNAs and the increased low-affinity ER in some leiomyoma, suggest that the redistribution of ER subtypes is associated with the pathological and/or normal growth of the myometrium.

Female↗
PubMed1997

A homeodomain point mutation of the Drosophila proboscipedia protein provokes eye loss independently of homeotic function.

The Drosophila homeotic gene proboscipedia (pb: HoxA2/B2 homolog) is required for adult mouthparts development. Ectopic PB protein expression from a transgenic Hsp70-pb minigene (HSPB) results in transformation of adult antennae to maxillary palps. In contrast, most tissues appear refractory to PB-induced effects. To study the basis of homeotic tissue specificity we are isolating and studying mutations that modify dominant HSPB-induced phenotypes. One HSPB point mutation (Arg5 of the homeodomain to His) removes homeotic activity in the mouthparts and antennae, but provokes a dose-sensitive eye loss. We show that eye loss can be induced by PB proteins that no longer effectively bind to DNA. The dose-sensitive eye loss thus appears to be mediated by specific, context-dependent protein-protein interactions.

Amino Acid Sequence↗
PubMed1997

Ras1-mediated modulation of Drosophila homeotic function in cell and segment identity.

Mutations of the Drosophila homeotic proboscipedia gene (pb, the Hox-A2/B2 homologue) provoke dose-sensitive defects. These were used to search for dose-sensitive dominant modifiers of pb function. Two identified interacting genes were the proto-oncogene Ras1 and its functional antagonist Gap1, prominent intermediaries in known signal transduction pathways. Ras1+ is a positive modifier of pb activity both in normal and ectopic cell contexts, while the Ras1-antagonist Gap1 has an opposite effect. A general role for Ras1 in homeotic function is likely, since Ras1+ activity also modulates functions of the homeotic loci Sex combs reduced and Ultrabithorax. Our data suggest that the modulation occurs by a mechanism independent of transcriptional control of the homeotic loci themselves, or of the Ras1/Gap1 genes. Taken together our data support a role for Ras1-mediated cell signaling in the homeotic control of segmental differentiation.

Animals↗
PubMed1997

Point mutations within and outside the homeodomain identify sequences required for proboscipedia homeotic function in Drosophila.

The Drosophila homeotic gene proboscipedia (pb) encodes a homeodomain protein homologous to vertebrate HoxA2/B2 required for adult mouthparts formation. A transgenic Hsp70-pb (HSPB) element that rescues pb mutations also induces the dominant transformation of antennae to maxillary palps. To identify sequences essential to PB protein function, we screened for EMS-induced HSPB mutations leading to phenotypic reversion of the HSPB transformation. Ten revertants harbor identified point mutations in HSPB coding sequences. The point mutations that remove all detectable phenotypes in vivo reside either within the homeodomain or, more unexpectedly, in evolutionarily nonconserved regions outside the homeodomain. Two independent homeodomain mutations that change the highly conserved Arginine-5 in the N-terminal hinge show effects on adult eye development, suggesting a previously unsuspected role for Arg5 in functional specificity. Three additional revertant mutations outside the homeodomain reduce but do not abolish PB+ activity, identifying protein elements that contribute quantitatively to pb function. This in vivo analysis shows that apart from the conserved motifs of PB, other elements throughout the protein make important contributions to homeotic function.

Amino Acid Sequence↗
PubMed1997

High polyunsaturated fatty acid, thromboxane A2, and alpha-fetoprotein concentrations at the human feto-maternal interface.

Polyunsaturated fatty acids (PUFA) like arachidonic (C20:4) and docosahexaenoic (C22:6) acids are essential for harmonious fetal development. This study evaluates, at near term, the distributions of free fatty acids (FFA) and their fetal carrier protein, alpha-fetoprotein (AFP) in the maternal (M) and fetal circulation (umbilical arteries (A) and vein (V)), focusing on the feto-material interface where maternal intervillous blood (I) contacts the fetal trophoblast. FFA concentrations in intervillous and maternal blood were similar, while those in umbilical arteries and vein were 2- to 4-fold lower (P < 0.001). There were more saturated FFA in umbilical vein (41%) and arteries (44%) blood than in maternal (30%) and intervillous (32%) blood (P < 0.001). Monounsaturated FFA predominated (P < 0.001) in maternal (43%) blood, but not in intervillous (35%), umbilical vein (33%) and arteries (31%) blood. Di-triunsaturated FFA were similar in intervillous and maternal (25%) blood and lower in umbilical vein and arteries (16%) (P < 0.001). PUFA were low in maternal (2.5%) blood and higher in intervillous and umbilical vein and arteries (9%, P < 0.001); consequently, C20:4 (40 microM) and C22:6 (16 microM) were the most abundant in the intervillous space. The AFP concentrations and AFP lectin-reactive isoforms were similar in intervillous and umbilical vein and arteries blood, but immuno-electrophoresis revealed a particular AFP conformation (less immuno-reactive, more anionic) in the intervillous space, suggesting that AFP is heavily loaded with PUFA at the feto-maternal interface. Prostacyclin derived from C20:4 was similar in all compartments but the thromboxane A2 concentration was 10-fold higher in intervillous blood than in maternal and umbilical vein and arteries blood. Thus the feto-maternal interface has a specific pattern of cell signalling molecules that might critically influence parturition.

Arachidonic Acids↗
PubMed1996

Interactions between phytoestrogens and human sex steroid binding protein.

The interactions of human Sex steroid binding protein (SBP) and the lignans [Nordihydrogaiaretic acid (NDGA) enterolactone (Ent), enterodiol (End)] and isoflavonoid phytoestrogens [Equol (Eq), diazein Dad), genistein (Gen)] were studied. The phytoestrogens had different dose-dependent inhibitory effects on steroid binding by SBP. Their relative efficiencies were: Ent> or = NDGA = Eq > Gen for displacing E2 and Eq > Ent > NDGA > Gen for displacing T. End and Dad were much less active. Scatchard analysis suggested that NDGA had similar non- competitive effects on T and E2 binding by reducing the number of binding sites without changing the association constants. But Eq seemed to inhibit E2 binding non-competitively and T binding competitively. NDGA binding to SBP reduced the immunorecognition of SBP by monospecific anti-SBP antibodies, suggesting that NDGA changed SBP immunoreactivity. Unlike NDGA, Eq binding to SBP caused no immunological changes in SBP, indicating qualitative differences in the effects of the lignan and isoflavonoid. Our results indicate that phytoestrogens may modulate the SBP activity and so influence the role of this protein in the delivery of hormonal information to sex steroid-dependent cells.

4-Butyrolactone↗
PubMed1995

Levels of homeotic protein function can determine developmental identity: evidence from low-level expression of the Drosophila homeotic gene proboscipedia under Hsp70 control.

The autonomous selector capacity of the homeotic proboscipedia (pb) gnee of the Drosophila Antennapedia Complex was tested. We introduced into the germline a P element containing a transcriptional fusion of a mini-gene for pb (normally required for formation of the labial and maxillary palps of the mouthparts) and the Hsp70 promoter. Uninduced expression of this Hsp70:pb element (HSPB) directs a novel, fully penetrant dominant transformation of antennae toward maxillary palps. Gene dosage experiments varying the number of HSPB elements indicate that the extent of the dominant transformation depends on the level of PB protein. At the same time, expression from the transgene also rescues recessive pb mutations. Finally, HSPB function may override the dominant antennal transformations caused by Antennapedia (Antp) mutations in a dose-sensitive manner, directing a switch of the antennal disc-derived appendage from ectopic leg to ectopic maxillary palp. This switch correlated with strikingly reduced ANTP protein accumulation when PB concentrations exceeded a genetically defined threshold level. These observations support a crucial role for quantitative aspects of pb function in determining segmental identity, including cross-regulatory events involved in this determination.

Animals↗
PubMed1995

Postprandial free fatty acids stimulate activity of human corticosteroid binding globulin.

The effect of postprandial variation of free fatty acids (FFA) on serum corticosteroid binding globulin (CBG) properties and cortisol (hydrocortisone) concentrations were explored in 11 women (20-30 yr) during 8 h after an oral load of tallow (26% C16:0, 18% C18:0, and 43% C18:1), oleic-sunflower (oleic-SF; 73% C18:1), sunflower (SF; 67% C18:2), and mixed oil (MO; 39% C18:1 and 48% C18:2). Serum FFA increased little after SF and MO but more than doubled in the late postprandial period (6 and 8 h) after oleic-SF (due to monounsaturated FFA) or tallow (due to saturated and monounsaturated FFA). CBG concentrations remained unchanged, but in relation with the postprandial elevation of serum FFA, CBG binding activity was increased after tallow or oleic-SF as a result of a combined two- to threefold increase in affinity constant and a 50% reduction in binding sites. Immunological and in vitro binding studies showed the changes in CBG behavior to be conformational and to be mediated mainly by monounsaturated FFA, especially C18:1. The modifications of CBG properties were associated with sustained high concentrations of cortisol (suppression of midday decrease) 6 and 8 h after tallow or oleic-SF. Thus dietary FFA may have an impact on bioavailability of glucocorticoids.

Adult↗
PubMed1994

In vivo effect of free fatty acids on the specific binding of glucocorticosteroids to corticosteroid binding globulin and liver receptors in immature rats.

Stimulating lipase activity with heparin (200 IU/kg b.w.) increased the plasma free fatty acid (FFA) concentration of immature rats (15 days). The effect of this elevated FFA concentration on glucocorticoid binding to corticosteroid binding globulin (CBG), and liver cytosol glucocorticoid receptor (GR), was analyzed. The plasma FFA concentration increased 2-fold, 10 minutes (P < 0.001), 20 minutes (P < 0.01), and 60 minutes (P < 0.01) post-heparin. The corticosterone (B) and progesterone concentrations were unchanged 60 minutes post-injection. The binding activity of immature rat CBG for B dropped 50% (P < 0.001) 60 minutes post-heparin injection, decreased B binding and increased plasma FFA were correlated (r = -0.8). The decreased B binding resulted from a 2-fold decrease in the apparent number of CBG binding sites; the affinity constant (Ka) remained unchanged. The liver cytosol endogenous FFA content of immature rats was also increased 2-fold, 60 minutes after heparin-induced lipolysis. The increased cytosol FFA, with no significant change in glucocorticoid, was accompanied by a significant decrease in dexamethasone binding to liver cytosol glucocorticoid receptor. The decrease resulted from a significantly lower apparent Ka for dexamethasone and fewer receptor binding sites (n). There was a good inverse correlation between Ka (r = -0.93) and n (r = -0.90) and the increased liver cytosol FFA content. Thus the higher plasma FFA induced in vivo by lipase activation or a standard FFA mixture probably causes conformational changes in CBG and GR, reducing glucocorticoid binding to immature rat CBG and liver GR.

Animals↗
PubMed1993

Free fatty acid-induced alterations in the steroid-binding properties of rat androgen-binding protein.

The free fatty acid (FFA) concentration in the epididymal cytosol of the adult rat was found to be 20-fold higher than in the serum. The binding of [3H] dihydrotestosterone to epididymal rat androgen binding protein (rABP) was modified by physiological concentrations of saturated and unsaturated fatty acids. Polyunsaturated fatty acids inhibited the binding more efficiently than monounsaturated or saturated fatty acids. Scatchard analysis and Dixon plots indicated that the number of binding sites decreased in presence of unsaturated fatty acids with an inhibition constant (Ki) of 4 microM for arachidonic acid (C20:4) and 20 microM for oleic acid (C18:1). These results indicate that unsaturated fatty acids induce alterations in rABP steroid-binding properties that could modulate the endocrine function of rABP.

Androgen-Binding Protein↗
PubMed1993

Stimulation of the binding properties of adult rat corticosteroid-binding globulin by a lipolysis-induced rise in plasma free fatty acids.

In vitro studies have shown that FFA induce conformational changes in human corticosteroid-binding globulin (CBG). We increased the plasma FFA concentrations of adult male rats by injecting heparin to determine whether such changes in CBG binding and immunological properties also occur in vivo. The in vivo transient activation of lipase by heparin produced a large increase in plasma FFA at 10 and 20 min (P < 0.01), which was maximal at 60 min (P < 0.005) and remained elevated at 120 min (P < 0.01) postinjection. This rise in FFA was associated with a 2- to 3-fold increase in the binding indices (C values; liters per g) of corticosterone (B) and progesterone to CBG 60-120 min postinjection (P < 0.001). There was a good positive correlation (r = 0.85) between the increase in B binding and the rise in plasma FFA in heparin-treated rats. The enhanced B binding to CBG resulted from a 2-fold increase in the apparent number of binding sites, without any significant change in the affinity constant (Ka). FFA extracted from postheparin plasma and a standard FFA mixture induced similar changes in B binding to purified mature rat CBG. The immunological behavior of CBG was not significantly changed after heparin-induced lipolysis, but the immunoreactivity of CBG from heparin-treated rats was more reduced by incubation with exogenous FFA than that from controls. FFA extracted from the plasma of heparin-treated rats and a standard FFA mixture both produced a dose-dependent drop in the immunodetection of pure CBG. These binding and immunological studies indicate that FFA mediate conformational changes in rat CBG in vivo. Thus, FFA, in addition to their roles as metabolic energy sources and components of complex lipids, can be rapid potent endogenous modulators of steroid-protein interactions.

Animals↗
PubMed1992

In vivo transient rise in plasma free fatty acids alters the functional properties of alpha-fetoprotein.

Previous in vitro studies have shown that unsaturated fatty acids (UFA) induce conformational changes in rodent and human alpha-fetoprotein (AFP). To determine whether such changes in the binding and immunological properties of rat AFP also occur in vivo, plasma free fatty acid (FFA) concentrations were increased in young male rats (15, 21 and 28 days old) by acute i.v. injection of heparin (200 IU/kg). Plasma estrogens (estrone and estradiol) did not change after injection of heparin. There was a large increase in plasma FFA 10-20 min post-heparin injection, with a return to normal 60 min later. This transient rise in FFA plasma was associated with a 50% drop (P less than 0.001) in the binding of estradiol to rat AFP of 15-, 21- and 28-day-old rats by reducing the number of binding sites (P less than 0.001), leaving the affinity constant (Ka) unchanged. FFA extracts from post-heparin plasma induced similar changes in estradiol binding to purified rat AFP. The rise in plasma FFA induced a loss of AFP immunoreactivity, in 21- (P less than 0.001) and 28-day-old rats (P less than 0.001), but not in 15-day-old rats. This age-dependent response correlated with the FFA/AFP molar ratio (38 in 15-day-old rats, 388 in 21-day-old rats, and 5600 in 28-day-old rats). These results indicate that an in vivo rise in FFA induces rapid and reversible conformational changes in AFP which may modulate the endocrine and immune function of this oncofetal protein.

Animals↗
PubMed1992

Alterations in the concentrations and binding properties of sex steroid binding protein and corticosteroid-binding globulin in HIV+patients.

The abnormal concentrations of steroid hormones and free fatty acids in the plasma of HIV-infected subjects are associated with qualitative and quantitative alterations in two of the major steroid hormones carrier proteins, sex steroid-binding protein (SBP) and corticosteroid-binding globulin (CBG). The properties of SBP and CBG in the sera of two age-matched groups of 67 men healthy blood donors (controls) and 64 HIV+subjects: 11 CDC group II and III (ASY), 6 CDC group IVA and 47 groups IV C1+D (AIDS) were analyzed. The HIV+patients had SBP concentrations 39-51% above those of controls. The sera of AIDS patients had higher SBP association constants (Ka) for testosterone than did those of the II, III and IVA groups and controls. In contrast, the CBG concentrations in all the HIV+subjects were similar to those of the controls. However, the binding properties of HIV+CBG were abnormal: the Ka's for cortisol and 17 alpha hydroxyprogesterone binding were 50% below normal, while the number of binding sites was significantly higher. Such changes in these carrier proteins could result from conformational transitions; they may cause abnormal transfer of hormonal information and/or steroid hormones metabolism, thus modifying the impact of steroids on the immune response in HIV+subjects.

17-alpha-Hydroxyprogesterone↗
PubMed1992

Effect of the aromatase inhibitor, 4 hydroxyandrostenedione, on the endotoxin-induced changes in steroid hormones in male rats.

The increase in circulating estrogen concentrations that follows injection of Escherichia coli endotoxin (Endo) may be due to increased aromatase activity. We have therefore analysed the effect of the aromatase inhibitor, 4 hydroxyandrostenedione (4OHA) on the steroid hormone response of male rats, particularly the dramatic increase in estrogens and decrease in androgens, induced by Endo. The concentrations of corticosterone (B), progesterone (P4), 17 alpha hydroxyprogesterone (17 alpha OHP4), androstenedione (delta 4), testosterone (T), estrone (E1) and estradiol (E2) were determined 2 hours after injection of increasing doses of 4OHA with and without Endo. The increase in serum estrogen concentrations and drop in serum androgen levels in response to Endo were blocked by a single dose of 4OHA. The effect of 4OHA appeared to be dose dependent. Low doses (30 mg/kg and 50 mg/kg) induced significant changes in the estrogen and androgen responses, but the high dose (100 mg/kg) blocked all changes in sex steroids induced by Endo. 4OHA did not alter the Endo-induced changes in other steroids.

Analysis of Variance↗
PubMed1992

Relationship between changes in serum estrone levels and outcome in human males with septic shock.

The effect of septic shock on the production of estrogens, other steroid hormones, and gonadotropins in men was investigated. Two groups of male patients in the early septic shock were studied over 3 days following their admission to the Intensive Care Unit. Group I (n = 9) patients recovered and group II (n = 6) patients died. The simplified acute physiological score was 13.5 +/- 1.5 for group I and 21.2 +/- 2.3 for group II (P less than .05). In group I patients, estrogen levels (particularly E1) were high on day 1 and decreased progressively (day 1: 3,515 +/- 884 pmol/L, day 2: 2,450 +/- 292 pmol/L, and day 3: 1,043 +/- 255 pmol/L). In group II patients, estrone levels were as high as in group I on day 1, but increased throughout the 3 days (day 1: 3,250 +/- 1,200 pmol/L, day 2: 4,495 +/- 930 pmol/L, and day 3: 6,123 +/- 966 pmol/L). There were few changes in gonadotropins and other steroid hormones, except that the testosterone levels were below normal in both patient groups, while cortisol was elevated in group II. The changes in serum E1 may provide an accurate marker of individual outcome.

Aged↗
PubMed1991

Phytoestrogens: new ligands for rat and human alpha-fetoprotein.

The binding of the lignans, enterolactone, enterodiol, nordihydroguaiaretic acid (NDGA), and the isoflavonic phytoestrogen equol, to human and rat alpha-fetoprotein (AFP) was studied. They had differential inhibitory effects (NDGA greater than equol greater than enterolactone greater than enterodiol) on the binding of estrone and estradiol to rat AFP and the binding of unsaturated fatty acid to both rat and human AFP. Inhibition was dose-dependent. The apparent dissociation constants (Kd) for phytoestrogens binding to AFP were: Kd NDGA = 5 +/- 1.2.10(-7) M, Kd equol = 6.7 +/- 0.8.10(-6) M, Kd enterolactone = 1.7 +/- 0.4.10(-5) M and Kd enterodiol = 2.2 +/- 0.6.10(-5) M. The Kd for estrone binding to rat AFP was increased by increasing concentrations of equol, but the number of esterone binding sites remained unchanged. This, plus the results of double-reciprocal plots, suggests that they compete for the same site(s). NDGA also competitively inhibited estrone binding at low NDGA concentrations (increased Kd), but high concentrations induced conformational changes in rat AFP, as both Kd and the number of binding sites (n) were altered. Both rat and human AFPs underwent changes in electrophoretic behaviour and loss of immunoreactivity with increasing NDGA, suggesting that NDGA binding induces conformational changes in the AFPs. However, equol did not alter the electrophoretic or immunological properties of either rat or human AFP, providing further evidence for qualitative differences in the effects of these diphenols. These findings indicate that phytoestrogens could play a role in AFP-dependent normal and pathological growth and development.

4-Butyrolactone↗