Search

The isotopic exchange reaction

We compare the cross sections for the transitions changing the projection of the total angular momentum of N2 +(2Σ) in collisions with 3He and 4He at very low collision energy. The fundamental states of the two nuclear spin isomers of N2 + are considered as well as the two fine structure levels of the first excited para level N=2. It is shown that the two fundamental states of the two nuclear spin isomers behave differently. For the fundamental para level N=0 of N2 +, the projection changing cross section is always negligible compared to the elastic one for both He isotopes. For the fundamental ortho level N=1 of N2 +, the spin-rotation interaction couples the different spin levels directly so the spin relaxation becomes a first order process. The associated resonances increase the projection changing cross section which remains smaller but becomes comparable with the elastic one. This is in contrast with the excited rotational levels of N2 +, which for the rotational deactivation and elastic channels are found to be equal around the resonances for the collisions involving 3He. These two channels are always larger than the projection changing one. We also find that, for transitions involving the fundamental rotational state, the domain of validity of the threshold laws discussed by Krems and Dalgarno [Phys. Rev. A 67, 050704 (2003)] for a potential decreasing faster than 1/r2 is shortened, due to the long range charge induced dipole potential. This effect is illustrated for the collisions of 3He with the fundamental para state of N2 +.

A comparative study of the inelastic scattering of 14N2 + and 14N2 in collision with 3He atoms is presented. The unrestricted nearside-farside (NF) method proposed by Connor [J. Chem. Phys. 104, 2297 (1995)] is applied to analyse the Close Coupling rotationally state selected angular distributions for four kinetic energies. These four energies illustrate different regimes of the dynamics. The relationships between the structures of the calculated differential cross-sections (DCS) and the different regions of the potential energy surfaces involved which can be extracted from semi classical models are here easily obtained from a simple reading of the (NF) figures. At the higher energy far-off the wells (1000 cm-1) the shape of the DCS are quite similar for the two systems and their nearside-farside components also, showing that the inelastic process is controlled by the short range repulsive part of the potential which is essentially the same for these two collisions. When the energy is decreased the differences between the two wells associated with the He–N2 + and He–N2 complexes are responsible for the differences between the DCS for the two systems. The farside component associated with the well become more and more prominent for the elastic scattering while inelastic scattering remains controlled by the repulsive core in a large angular interval. The nearside farside analysis gives also a new picture of a resonance which is regarded as an equilibrium between the repulsive and the attractive parts of the potential.

International audience; For several decades, global change has resulted in an increase in runoff in the Sahelian belt, provoking major changes in the quality and quantity of sediments transported by drainage networks. One of the astonishing consequences is the establishment of numerous permanent lakes. The origins of particulate organic matter (OM) preserved within lacustrine sediments of three lakes were investigated by coupling optical observations (palynofacies) and bulk geochemistry (Rock-Eval 6 pyrolysis). An initial estimate of particulate organic carbon (OC) stored in these lacustrine sediments was assessed. Soil organic matter (SOM) was sampled from the surface (0-10 cm) of various land-use and land-cover areas and was characterized and compared with sedimentary organic matter. Our results reveal that Lake Tankalawal is subjected to high autochthonous organic sedimentation (TOC ranges between 3.0 and 10.0 wt.%), while lakes Bangou Kirey and Bi are characterized by weak sedimentation of non-indigenous OM originating from the soil erosion and shore vegetation (TOC < 3.0 wt.%). In sediments, the effects of early diagenesis on the OM induce not only a loss of labile and aquatic OM but also a significant loss of terrestrial OM, which is supposed to be more resistant than its aquatic counterpart. Both the preservation of OM in top sediments and the relative preservation of terrestrial OM impact the OC storage in lakes. Indeed, OC storage in lacustrine sediments (Lake Bangou Kirey) was two to seven times higher than the OC storage in Sahelian soils, where greater contributions of terrestrial OM to sedimentary OM are associated with lower OC storage in lacustrine sediments. For lakes Bangou Kirey and Bi, OC accumulation rates were also assessed; due to the identification of a sedimentary limit corresponding to the establishment of permanent lakes dated earlier 1960s. High OC fluxes were estimated and ranged between 104 and 213 g OC m− 2 yr− 1. Compared with other OC accumulation rates for various African lakes, these high values are similar to those calculated for reservoirs and are related to anthropogenic pressure, soil textures favoring erosion, and proper physical and chemical conditions for OM preservation in sediments. Accordingly, in these Sahelian environments that are generally viewed as non-efficient in storing OC, we claim that global change could promote a new OC sink. If other similar studies reinforce our assertion, then regional C budgets should be revisited.

The hydrodynamic, engineering, and chromatographic variables affecting scale-up of countercurrent chromatography (CCC) are examined. The predictable and linear scale-up from the current laboratory scale technology to industrial process scale, capable of kgm/month in the first phase, is demonstrated. Continued research will prepare the way for a new generation of tonne/annum capacity high throughput, high resolution CCC machines for pilot and plant scale separations of a range of bioprocess products.

This study describes how scale-up in countercurrent chromatography (CCC) can be simply predicted on a process scale CCC device by running a preliminary analytical-sized sample and having knowledge of the stationary-phase retention at scale-up conditions. Results have shown that simple experimentation can lead within a day to a process with the capability of several kilograms per day (tons per year) compound yield, and that this is feasible with benchtop CCC units.

We study the effect of a magnetic field on the spin-depolarization cross sections of $\text{N}_{2}{}^{+}(^{2}\ensuremath{\Sigma})$ in collisions with $^{3}\text{H}\text{e}$ and $^{4}\text{H}\text{e}$ at very low collision energy. The fundamental states of the two nuclear spin isomers of $\text{N}_{2}{}^{+}$ are shown to respond quite differently to the application of the field. When the applied magnetic filed increases, the spin-depolarization cross sections are found to monotonously decrease for the fundamental paralevel $N=0$ of $\text{N}_{2}{}^{+}$ whereas they monotonously increase for the fundamental ortholevel $N=1$ of $\text{N}_{2}{}^{+}$. This effect is found to be amplified around the resonances and an explanation based on the different mechanism of spin flipping acting for these two levels is proposed. We discuss the sensitivity of the results on the change of relative mass when $^{3}\text{H}\text{e}$ is replaced by $^{4}\text{H}\text{e}$. We also explore the tuning of the inelastic cross sections by examining the variation of the scattering length as a function of the magnetic field and locate several narrow Feshbach resonances.

In a recent work we compared spin-free collisions of N{sub 2}{sup +} with {sup 3}He and {sup 4}He using a newly developed potential energy surface. We found that such collisions exhibit a strong isotope effect in the ultracold regime. In the present study we consider the role of the spin-rotation interaction in cold He-N{sub 2}{sup +} collisions. We find that our spin-free vibrational quenching cross sections are not affected. Conversely, the cross sections for rotational quenching are strongly modified in the ultracold regime and exhibit resonance structures which are analyzed. These structures, which are not observed when the spin-rotation interaction is omitted, are shown to be due to the Feshbach resonances associated with the nonconservation of parity.

the differentiation of NG108−15 neuroblastoma cells and rat striatal neurons in primary culture, assessed by neurite outgrowth. Expression of voltage-gated Ca2+ channels was also analysed in NG108−15 cells using Fura2 imaging. The unpaired Student’s t-test and one-way ANOVA followed by Dunnett’s test were performed (using the Prism software) for two-sample and multiple comparisons, respectively. Expressing 5-HT6 receptors in NG108−15 neuroblastoma triggered neurite outgrowth and induced expression of functional voltage-gated Ca2+ channels. These effects were not further enhanced by an agonist (WAY181,187, 1mM) but were prevented by SB258,585 (10mM), a selective 5-HT6 antagonist. Thus, upon SB258,585 treatment, cells showed a decrease in neurite length of 48.4% compared to untreated cells (22.5±1.5 mm vs. 43.5±1mm, n = 4, p< 0.0001). Expression of a dominantnegative (DN) Cdk5 or treating cells with roscovitine (pharmacological inhibitor of Cdk5) likewise inhibited NG108−15 cells differentiation induced by 5-HT6 receptor expression (control: 69.8±3.6mm; DN Cdk5: 42.8±3mm, p< 0.0001; roscovitine: 28.1±1.5mm, n = 3, p< 0.0001). SB258,585 also impaired the association of 5-HT6 receptor with Cdk5 in NG108−15 cells, as determined by co-immunoprecipitation, suggesting that this interaction was necessary for induction of differentiation. Treating striatal neurons with either SB258,585 or roscovitine immediately after seeding decreased neurite length, as determined 24 hrs later (untreated: 29.9±0.7mm; SB258,585: 23.7±0.6mm, p< 0.0001; roscovitine: 20.2±1.0mm, n = 4, p< 0.0001). Conversely, exposure of neurons to WAY181,187 did not significantly affect neurite outgrowth (p = 0.43). Further supporting a role of endogenously expressed receptors in differentiation of striatal neurons, silencing 5-HT6 receptor expression in cultured neurons significantly reduced neurite length (control siRNA: 29.6±0.8mm, 5-HT6 receptor siRNA: 17.2±0.9mm, n = 3, p< 0.0001). Conclusions: Complementing work indicating that 5-HT6 receptors modulate neuronal migration [3], the present data show that 5-HT6 receptors developmentally promote neuronal differentiation and reveal a critical role for Cdk5 in this process. These novel insights into molecular substrates underlying neurodevelopmental effects of 5-HT6 receptors are of potential importance to the pathophysiology and treatment of early-onset CNS conditions like autism-spectrum disorder and schizophrenia.

[3H]-SR 121463 is the first radiolabeled selective nonpeptide vasopressin V2 receptor antagonist ligand that has been reported to date. In the present work, we studied the binding properties of [3H]-SR 121463 for renal V2 receptors from animal and human origins.Binding studies were performed with [3H]-SR 121463 in Chinese hamster ovary (CHO) cells transfected with the human V2 receptor and in various kidney preparations expressing the native V2 receptors (rat, rabbit, dog, pig, monkey, and human). Autoradiographies were performed in rat and human kidney sections.[3H]-SR 121463 binding to CHO cells stably transfected with the cloned human renal V2 receptor was specific, highly stable, time dependent, saturable, and reversible. A single population of high-affinity binding sites was identified (Kd = 0.94 +/- 0.34 nmol/L, Bmax = 9876 +/- 317 fmol/mg protein). Of note, [3H]-SR 121463 revealed a higher number (about 40%) of V2 sites than [3H]-AVP in the same preparation. Displacement of [3H]-SR 121463 binding by reference peptide and nonpeptide vasopressin/oxytocin compounds exhibited a typical AVP V2 profile. [3H]-SR 121463 also displayed a high affinity for native V2 receptors in several kidney preparations from rat, pig, dog, rabbit, bovine, monkey, and human. The autoradiographic experiments using rat and human kidney sections showed intense labeling in the medullopapillary region and lower intensity in the cortex, consistent with a main localization of V2 receptors on collecting tubules.[3H]-SR 121463 is a useful ligand for the specific labeling of animal and human V2 receptors and could be a suitable probe for the search and in situ localization of V2 sites.

We assessed the role of vasopressin (VP) for the hemodynamic response to pneumoperitoneum in pigs. Four groups of anesthetized pigs were investigated. Nine pigs were intraabdominally insufflated with CO2 and eight were intraabdominally insufflated with argon; eight pigs received an IV injection of 1 mg/kg SR 49059, a VP antagonist, before CO2 insufflation; and six pigs received SR 49059 alone. Hemodynamics, plasma concentrations of VP and vasoactive hormones, and Paco2 were measured. Data were analyzed by using analysis of variance, Student’s t-test, and Mann-Whitney U-test. Five minutes after insufflation, changes in systemic vascular resistance (SVR) were significantly correlated with changes in VP (r 5 0.72; P 5 0.005) but not with changes in epinephrine, norepinephrine, renin activity, or Paco2. SVR increased during CO2 insufflation but not during argon insufflation or CO2 insufflation with a preceding infusion of SR 49059. The SR 49059 injection itself resulted in increases in heart rate and cardiac output and decreases in blood pressure and SVR. We conclude that, during CO2 pneumoperitoneum in pigs, absorbed CO2 initiates a pathophysiological process that stimulates VP release. Hence, VP most likely plays a key role in the hemodynamic response to a CO2induced pneumoperitoneum. Implications: Intraabdominal insufflation of CO2 is associated with hemodynamic and hormonal changes. Investigating CO2 and argon-insufflated pigs and using a vasopressin antagonist, we found that CO2 insufflation released vasopressin, which, in turn, induced hemodynamic perturbances. (Anesth Analg 1999;89:278‐83)

The nature of vasopressin (VP) receptors present in normal and tumoral human adrenal was investigated using various experimental approaches. Specific VP-binding sites were detected by autoradiography using [3H]arginine VP as a radioligand in adrenal cortex and medulla. The V1a receptor subtype was expressed in the two parts of the gland, as shown by pharmacological studies and RT-PCR experiments. By contrast, the V1b receptor subtype was only expressed in medullary chromaffin cells. This was confirmed by the characterization of V1b transcripts detected in adrenal medulla tissues. In pheochromocytoma, we also detected functional V1b receptors. These receptors triggered intracellular calcium mobilization from intracellular pools and were involved in catecholamine secretion. Binding experiments performed on pheochromocytoma plasma membrane preparations also revealed V1a vasopressin-binding sites, whose roles and cellular localization have not yet been determined. RT-PCR experiments confirmed these data; 100% and 80% of the five tumors tested exhibited V1a and V1b transcripts, respectively. Perifusion experiments also demonstrated that some pheochromocytomas may secrete large amounts of VP. Our findings imply that VP locally secreted by human adrenal medulla may regulate adrenal function by acting on V1a or V1b receptors. More interestingly, we demonstrate that one pheochromocytoma oversecretes VP. In this particular case, this may contribute to the increase in blood pressure observed.

In mammals, vasopressin is known to be synthesized in the hypothalamus and released in the blood stream at the pituitary level. This neuropeptide is also synthesized and secreted by the adrenal medulla in many species including human. Moreover, agents like acetylcholine and corticotropin releasing factor stimulates its basal secretion. V1a vasopressin receptors are present in the adrenal cortex and are involved in steroids secretion (aldosterone in the zona glomerulosa and glucocorticoids in the zona fasciculata of some species). These receptors are coupled to phospholipase C beta and to dihydropyridine-sensitive calcium channels via heterotrimeric G proteins differing by their sensitivities to pertussis toxin. The adrenal medulla, from many species, exhibits V1a vasopressin receptors. In rat adrenal medulla, functional V1b vasopressin receptors could also be characterized. These receptors stimulate catecholamines secretion via activation of phospholipase C beta and subsequent mobilization of intracellular calcium. The adrenal medulla secretes AVP and exhibits functional vasopressin receptors. The adrenal cortex also possesses functional vasopressin receptors and is in contact with adrenal medulla via "medullary rays". We may thus reasonably conclude that AVP physiologically regulates adrenal gland functions via autocrine/paracrine mechanisms.

Our recent studies have shown that regulation of uterine oxytocin (OT) binding involves at least two different mechanism: Estradiol (E2)-induced upregulation is accompanied by an increase in OT receptor (OTR) mRNA accumulation, implying that the E2 effect is mediated via increased OTR gene transcription and/or OTR mRNA stabilization. In contrast, P (P)-induced OTR down-regulation occurs via a novel non-genomic mechanism, involving a direct interaction of P with the OTR at the level of the cell membrane. We found that P specifically binds to the OTR and inhibits its ligand binding and signalling functions. Physiological levels of P repress in vitro the ligand binding capacity (Bmax) of the OTR by50%. When expressed in CHO cells, the OTR provides a high affinity (Kd: 20nM) membrane binding site for P. OT-induced inositol phosphate production and intracellular calcium mobilization is inhibited 85% and 90%, respectively, by P. These effects are specific as signalling and binding functions of the closely related V1a vasopressin receptor remain unaffected by P, and as other, related steroids are devoid of any effect on OTR binding or signalling functions. The present observation of a specific interaction of a steroid with a G-protein-linked receptor defines a new mechanism of non-genomic steroid action and uncovers a novel level of crosstalk between steroid and peptide hormone action.

The hormone-mediated intercellular Ca2+ waves were analyzed in multiplets of rat hepatocytes by video imaging of fura2 fluorescence. These multicellular systems are composed of groups of several cells (doublets to quintuplets) issued from the liver cell plate, a one cell-thick cord of about 20 hepatocytes long between portal and centrolobular veins. When the multiplets were homogeneously bathed with the glycogenolytic agonists vasopressin, noradrenaline, angiotensin II and ATP, they showed highly organized Ca2+ signals. Surprisingly, for a given agonist, the primary rises in intracellular Ca2+ concentration ([Ca2+]i) originated invariably in the same hepatocyte, then was propagated in a sequential manner to the nearest connected cells (cell 2, then 3, cell 4 in a quadruplet, for example). The sequential activation of the cells appeared to be an intrinsic property of multiplets of rat hepatocytes. The same sequence was observed at each train of oscillations occurring between cells. The order of [Ca2+]i responses was modified neither by repeated additions of hormones nor by the hormonal dose. The mechanical disruption of an intermediate cell did not prevent the activation of the next cell. These results suggest that each hepatocyte in the multiplet displays its own sensitivity to the hormone and that a gradient of sensitivity between each cell could be responsible for directing the intercellular Ca2+ wave. To test this hypothesis, we selectively isolated rat hepatocytes from periportal (PP) and perivenous (PV) areas of the liver cell plate. Periportal (PP) and perivenous (PV) rat hepatocyte suspensions were loaded with quin2/AM and hormonal responses were studied in a spectrofluorimeter. Noradrenaline, angiotensin II, and vasopressin-induced [Ca2+]i rises were greater in PV than in PP hepatocytes. In contrast, PP cells were more responsive than PV cells to ATP. The function of the InsP3 receptor (InsP3R) was also studied by measuring the InsP3-mediated 45Ca2+ release from permeabilized PP and PV hepatocytes. In permeabilized PP and PV hepatocytes, internal Ca2+ stores displayed the same loading-kinetics, the responses to InsP3 were similar, and the sizes of InsP3-sensitive compartment were not different. In a further study, we investigated by video microscopy in fura2-loaded multicellular systems of rat hepatocytes, the mechanisms controlling intercellular propagation of the Ca2+ wave and coordination of Ca2+ signals induced by the different hormones. Using focal microperfusion which allows local perfusion of any cell of the multiplet, rapid agonist removal during the Ca2+ response and microinjection, we found that second messengers and [Ca2+]i rises in one hepatocyte cannot trigger Ca2+ responses in connected adjacent cells, suggesting that diffusion across gap junctions, while required for coordination, is not sufficient by itself for the propagation of the intercellular Ca2+ wave. In addition, focal microperfusion and intermediate cell disruption experiments revealed very fine functional differences (hormonal delay, frequency of [Ca2+]i oscillations) between hormone-induced Ca2+ signals, even between two adjacent connected hepatocytes. Recent unpublished results performed in suspensions of PP and PV rat hepatocytes supported the view of a major role played by vasopressin receptors (V1a) in genesis and orientation of the Ca2+ wave. Vasopressin binding sites, V1a mRNAs detected by RNAse Protection Assay, and vasopressin-induced InsP3 production, were more abundant in PV than in PP cells. A gradient of hormone receptors could orientate the propagation of the Ca2+ wave in multicellular systems and in liver cell plate. These results suggest that the intercellular Ca2+ wave in multicellular systems of rat hepatocytes is propagated through mechanisms involving at least three factors. (ABSTRACT TRUNCATED)

In 3-day primary cultures of rat glomerulosa cells, a 30-min pre-incubation with either 10 microM colchicine (a microtubule-disrupting agent) or 10 microM cytochalasin B (a microfilament-disrupting agent) decreased angiotensin II (Ang II)-induced inositol phosphate accumulation by 50%. Moreover, both drugs decreased inositol phosphate production induced by fluoroaluminate (a nonspecific activator of all G proteins), indicating that both microtubules and microfilaments are essential for phospholipase C activation. Analysis of microfilament- and microtubule-enriched fractions and immunoprecipitation of actin and tubulin revealed that the alpha(q)/alpha11-subunit of the G(q/11) protein was associated with both structures. Ang II stimulation induced a rapid translocation of alpha(q)/alpha11, microfilaments, and microtubules to the membrane and induced a time-dependent increase in the level of alpha(q)/alpha11 associated with both microfilaments and microtubules. Moreover, double immunofluorescence staining clearly showed a colocalization of the alpha(q)/alpha11-subunit of the G(q/11) coupling protein and microfilament distribution. These associations and plasma membrane redistribution under Ang II stimulation indicate that microfilaments and microtubules are both involved in phospholipase C activation and inositol phosphate production. Moreover, our results indicate that the alpha(q)/alpha11 protein is closely associated with cytoskeletal elements and is found both at the plasma membrane level as well as on intracellular stress fibers.

In rat glomerulosa cells, vasopressin stimulates intracellular calcium mobilization via at least two distinct mechanisms: the release of calcium from inositol-1,4,5-P3-sensitive stores and the activation of transmembrane calcium influx. In this study, we focused on the second mechanism through three experimental approaches. By videomicroscopically examining Fura-2-loaded cells, we demonstrate that vasopressin induces a dose-dependent and receptor-mediated calcium influx fully inhibited by either 1 microM nifedipine or a pertussis toxin pretreatment and potentiated by 1 microM BAY K 8644. Patch-clamp experiments also indicate that vasopressin stimulates L-type calcium current by 87% and only weakly inhibits T-type calcium current. To further characterize the coupling between the vasopressin receptor and the dihydropyridine calcium channel, we performed binding studies using tritiated nitrendipine. With this technique, we showed that on intact cells, vasopressin is able to increase the specific binding of tritiated nitrendipine in a dose-dependent manner (Kact = 2 nM). Pharmacological studies using a series of vasopressin analogs revealed that this effect is mediated via a V1a vasopressin receptor subtype. Furthermore, the vasopressin-stimulated nitrendipine binding was sensitive to pertussis toxin pretreatment, which affected only the maximum binding capacity of nitrendipine-binding sites. More interestingly, we demonstrate that vasopressin still increases nitrendipine binding to plasma membrane preparation and that GTP is absolutely necessary for such a hormonal effect. Altogether, these data confirm the existence of a tight and direct coupling between the V1a vasopressin receptor and a dihydropyridine calcium channel via a pertussis toxin-sensitive G protein.

Ionic currents of primary cultured glomerulosa cells from human adrenal glands were studied with the patch-clamp technique. Two types of outward K+ currents and two types of inward Ca2+ currents were described. The transient outward K+ current activated at potential positive to -40 mV and demonstrated a marked time-dependent inactivation. It was blocked by 4-aminopyridine but not tetraethylammonium. A second type of outward current activated rapidly at the depolarization onset and then increased slowly with no time-dependent inactivation. The transient inward T-type Ca2+ current was activated for potential positive to -60 mV with a maximal current amplitude at -30 mV and zero current voltage at +40 mV; it was completely inactivated for membrane potential positive to -40 mV. The pharmacological studies of the T-type channel showed that Ni2+ was a potent blocker but that the channel was not sensitive to dihydropyridine. The long-lasting inward Ca2+ current was activated for potentials positive to -20 mV with a maximum current amplitude at +70 mV. This current was increased by the agonist Bay K 8644 and blocked by the antagonist nifedipine; in addition, it was blocked by Cd2+ but less sensitive to Ni2+. This study revealed that glomerulosa cells from human adrenal demonstrated the presence of K+ and Ca2+ currents similar to those found in rat and bovine cells. Moreover, the main stimuli of aldosterone secretion, ACTH and angiotensin II, induce an increase in aldosterone secretion which is inhibited in a Ca(2+)-free external medium.

Cholinergic effects on kidney function have been observed in some mammals but the intrarenal localization and the cellular mechanisms of these effects are poorly defined to date. The aim of this work was to study the effects of carbachol on phosphoinositide metabolism in freshly isolated rat glomeruli labeled with myo-[3H]inositol. Carbachol rapidly and markedly stimulates phosphoinositide metabolism with a 50% effective concentration of 3 microM. The enormous magnitude of the response is enlightened by the use of 10 mM lithium, which provokes in the presence of the agonist a large accumulation of inositol phosphates and a corresponding depletion of cellular free inositol. The response is inhibited by 85% by pirenzepine, is pertussis toxin insensitive, and shows no desensitization at maximum dose of carbachol up to 40 min of stimulation.

We give a brief account of the effect of a magnetic field on the spin-depolarization cross sections of N2 +(2Σ) in collisions with 3He at very low collision energy. The fundamental states of the two nuclear spin isomers of N2 + are shown to respond quite differently to the application of the field. When the applied magnetic field increases, the spin depolarization cross sections are found to monotonously decrease for the fundamental para level N=0 of N2 +, whereas they monotonously increase for the fundamental ortho level N=1 of N2 +. This effect is found to be amplified around the resonances and an explanation based on the different mechanism of spin flipping acting for these two levels is proposed. We also explore the tuning of zero energy Feshbach resonances by examining the variation of the scattering length as a function of the magnetic field. We use the Smith Q matrix to calculate the lifetimes of the complexes associated with these resonances and discuss the law of variation as a function of the magnetic field of the Q matrix eigenvalues across such resonances.

Numerous hormones or neurotransmitters regulate the activity of their target cell via the activation of a specific phospholipase C. Two intracellular second messengers are generated, diacylglycerol which activates protein kinase C and inositol (1,4,5)P3 which mobilizes calcium from intracellular stores. Both these molecules trigger within the cell biological effects associated with the hormone considered. In these transduction mechanisms, calcium mobilization is a consequence of primary activation of phospholipase C. However calcium is also able to regulate phospholipase C activity since, in many cellular systems, calcium by itself stimulates phosphoinositol lipid metabolism. Such results imply that all molecules which modify the intracellular calcium concentration may be considered as potent regulators of phospholipase C activity. Data obtained on different cell systems favours this hypothesis: Activators of calcium influx stimulate intracellular inositol phosphate accumulation and molecules which reduce intracellular calcium concentration reduce inositol phosphate production. Thus calcium may play an important role in the regulation of hormonal sensitive phospholipase C activities.

The first quantum mechanical investigation of the rotational deactivation of HF induced by collisions with ortho- and para-H(2) molecules is reported. Ab initio potential energy calculations are carried out at the coupled cluster level with single and double excitations, using a quadruple-zeta basis set. The global rigid rotor four-dimensional potential energy surface is obtained by fitting ab initio points with a least squares procedure for the angular terms and interpolating the radial coefficients with cubic splines. It is shown that the equilibrium structure of the H(2)-HF complex is T-shaped and the well depth is found to be 359 cm(-1). Close coupling scattering calculations are performed at collision energy ranging from 10(-2) to 1600 cm(-1). A comparison of the rotational quenching of HF with para-H(2) and (4)He is used to validate our potential energy surface. The rotational quenching cross sections of HF by ortho- and para-H(2) are also compared and found to be very different. An explanation of these differences based on a resonance mechanism is proposed.

Cells from the zona glomerulosa of rat adrenals were isolated and maintained for 3 days in primary culture. Specific vasopressin binding was determined by using [3H]vasopressin. [3H]Vasopressin binding was time-dependent (half-time of about 2 min for 6 nM free ligand) and reversible on addition of unlabelled vasopressin (80% dissociation within 30 min). Dose-dependent [3H]vasopressin binding at equilibrium indicated that vasopressin interacted with two populations of sites: high-affinity sites (dissociation constant, Kd = 1.8 nM; maximal binding capacity = 10 fmol/10(6) cells) and low-affinity sites. Vasopressin increased the cellular content of labelled inositol mono-, bis- and tris-phosphate in cells prelabelled with myo-[3H]inositol. The vasopressin concentration eliciting half-maximal inositol phosphate accumulation was very close to the Kd value for vasopressin binding to high-affinity sites. Competition experiments using agonists and antagonists with enhanced selectivity for previously characterized vasopressin receptors indicated that vasopressin receptors from rat glomerulosa cells are V1 receptors of the vascular or hepatic subtype. The detected specific vasopressin-binding sites might represent the specific receptors mediating the mitogenic and steroidogenic effects of vasopressin on glomerulosa cells from rat adrenals.

By means of scanning electron-microscopic investigations, thorn-like projections (TLP) were observed in the cilio-secretory epithelium of the ventral surface of the human endo-cervix. These projections, which seem to be characteristic of a new cell type, were seen at different stages of the menstrual cycle (days 8, 14 and 21) in 4 apparently healthy fertile women. Striking differences in length, with a maximum size at midcycle, suggest an evolutive process throughout the menstrual cycle. The origin and possible physiological role of TLP in the reproductive process are discussed.

Multiple forms of acetylcholinesterase have been studied in Goldfish adapted to 15 degrees C and 25 degrees C. Three forms of enzyme (sedimenting at 18S, 12S and 5S) could be distinguished by sucrose gradient centrifugation. Their molecular and enzymatic properties did not vary with environmental temperature. It was noted that the relative proportion of the three forms differed between brain and muscle extracts, the 18S forms predominating in muscle (about 75 per cent) the 12S forms being more abundant in the brain, suggesting a physiological differentiation between these forms. The latter form varied from 48 per cent to 70 per cent in Goldfish adapted to 15 degrees and 25 degrees C respectively. The relative efficiency of saline buffer solubilization for each form also depended on the tissue used and in all cases decreased with rising environmental temperature. These two facts are taken to reflect interactions between acetylcholinesterase and membrane that depend on the nature of the membrane and probably involve the aliphatic chains of its phospholipids. The greater case of solubilization found in the low temperature adapted fish may be a function of the increased fluidity of the cellular membranes formed at lower temperatures.

The anatomy of the ventral epithelial surface of the human uterine endocervix was investigated by S.E.M. from biopsies taken at various periods of the menstrual cycle and from whole cervices obtained by hysterectomy on the 17th and 22nd day. Ciliated and secretory cells which constitute the epithelial surface appeared to be associated at random in various manners without apparent connection with the menstrual cycle. Considerable variation was observed at the same cycle period, depending on the patients. The release of mucus took place mainly according the macro-apocrine extrusion process, whose evolution was clear throughout menstrual cycle. Apocrine extrusions were found to be particularly intense and numerous from the 12th to the 16th day, with a peak on the 14th day. Some ciliated cells were also seen to exhibit protuberant mucus protrusion. The merocrine extrusion process was seen very rarely and thus seemed to be of minor importance. An evolutive provess also seems to occur at the level of ciliated cells: from the 9th to the 16th day, some secretory cells were seen to bear miniciliae of increasing length beside their typical microvillae. This could be interpreted as a transformation process of secretory into ciliated cells during the second third of the menstrual cycle.

The ligand specificity of rat adenohypophyseal vasopressin receptors was directly compared to that of peripheral receptors of the V1 and V2 types. For this purpose a series of 15 recently designed vasopressin antagonists was used. The affinities of these antagonists for rat adenohypophyseal membranes were deduced from the determination of the concentration-dependent inhibition of [3H]vasopressin binding. In parallel experiments the corticotropin (or anti-corticotropin)-releasing activities of the tested peptides were determined on freshly dispersed rat adenohypophyseal cells. All peptides tested which were found to be antagonists of the vasopressor and antidiuretic responses to vasopressin in vivo behaved as antagonists of vasopressin-induced corticotropin release. There was a close correlation between the relative affinities of the analogues tested for binding to adenohypophyseal membranes and their relative potencies in inhibiting vasopressin-induced corticotropin release, indicating that the detected vasopressin-binding sites are the receptors involved in the vasopressin effect on corticotropin secretion. No correlation could be demonstrated between anti-corticotropin-releasing activities and either anti-antidiuretic or antivasopressor potencies of the antagonists tested. A direct comparison of the ligand specificities of adenohypophyseal receptors on the one hand, and V1 (hepatic) and V2 (renal) receptors on the other hand, showed that most of the antagonists discriminated very efficiently between adenohypophyseal and either hepatic or renal receptors. The selectivity index reaches values as high as 260,000 for desGly(NH2)9 [1-(beta-mercapto-beta, beta-cyclopentamethylenepropionic acid), 2-D-O-ethyl-tyrosine, 4-valine] arginine vasopressin. It is concluded that adenohypophyseal receptors represent a novel type of vasopressin receptors. Based on the observation that adenohypophyseal receptors, like hepatic or vascular V1 receptors, do not appear to be coupled to adenylate cyclase, we propose that adenohypophyseal receptors could be designated as V1b receptors as opposed to the V1a receptors previously characterized on liver and blood vessels.

As previously reported, it is possible to solubilize vasopressin-receptor complexes formed in rat kidney membranes by the use of Triton X-100. Ultracentrifugation on sucrose gradients and elution through molecular sieving columns of these soluble extracts revealed the existence of two molecular forms of vasopressin-receptor complexes (molecular weight = 200,000 and 100,000, respectively). These two molecular forms of vasopressin-receptor complexes can be partially purified and exhibit different properties: (a) The light form is more sensitive to thermal dissociation than is the heavy form. (b) The presence of guanyl nucleotide affects the dissociation rate of only the heavy form of the vasopressin-receptor complex. (c) The light form seems to be convertible to the heavy form by increasing the duration of incubation between the membranes and the tritiated hormone. (d) Guanyl nucleotides affect the distribution of the two molecular forms of the receptor (decrease of the relative amount of the heavy form). These data provide evidence for interaction between vasopressin-receptor complexes (light form) and another protein component, which may be a GTP-binding protein.

Systematic measurements of basal blood prolactin (PLR) levels were conducted in 383 patients with sperm anomalies or features of marital sterility of hyposterility, and 47 subjects consulting for sexual disorders. PRL levels were rarely abnormal in the patients in the two groups. Though systematic measurement of PRL levels should still be continued in patients with sexual disorders, the author refutes their usefulness in subjects consulting for sterility, their value in these cases being determined after the consultation and clinical examination, and, in some cases, by examination of the sperm.

The exposure of WRK1 cells to arginine vasopressin (AVP), lysine vasopressin, or oxytocin for 18 h at 37 degrees C induced a homologous desensitization of the vasopressin- (VP) receptors. Dose-response curves of [3H]lysine vasopressin binding to control and desensitized WRK1 cells revealed a decrease in the maximal number of binding sites without any modification of its affinity (Kd values = 4.40 +/- 0.76 nM and 4.65 +/- 0.78 nM for control and desensitized conditions, respectively). The phenomenon was time- and dose-dependent. It was directly related to receptor occupancy, since the concentration of VP analogues leading to a half-maximal occupancy of VP receptors was closely related to the concentration of the corresponding analogue leading to a half-maximal decrease in VP-binding sites. It was also agonist-specific, since the V1 vasopressin antagonist desGly9d(CH2)5[D-Tyr(Et)2]VAVP was unable to affect the number of receptors. These desensitization processes were completely inhibited when the functional coated pits present in WRK1 cells were suppressed, indicating that the loss of VP-binding sites was related to receptor internalization. The exposure of WRK1 cells to a vasopressin agonist for 18 h also led to an inhibition of the vasopressin-sensitive phospholipase C activity. It was time- and agonist-dose-dependent, and occurred without any detectable changes in apparent affinity values (1.40 +/- 0.04 and 1.90 +/- 0.36 nM for control and desensitized cells, respectively). Control experiments showed that these inhibitions could not have been caused by a decrease in the labeling of inositol lipids. It is likely that they were mainly due to receptor internalization since (i) the hormonal treatment did not modify the basal level of phospholipase C; (ii) the maximal loss of VP-binding site was similar to the maximal inhibition of VP-stimulated IP accumulation; (iii) the recoveries of both VP-binding sites and VP-sensitive phospholipase C activity followed exactly the same time course (t1/2 = 4 h). In addition to this homologous desensitization of VP-sensitive phospholipase C activity, AVP also induced heterologous desensitization of bradykinin-sensitive phospholipase C activity. However, this effect was relatively weak (maximal inhibition 17 +/- 3%). The time course of VP-sensitive phospholipase C desensitization was more rapid than that of VP-receptors, indicating that desensitization involved at least two distinct steps, a rapid uncoupling step, and a later loss of vasopressin receptors.(ABSTRACT TRUNCATED AT 400 WORDS)

5-Hydroxytryptamine (5-HT) stimulates the accumulation of inositol-trisphosphate in WRK1 cells, a cell line originating from a rat mammary tumor. 5-HT acts via a single receptor type for which it has an affinity constant estimated to be 1.27 microM. A series of agonists known to act at 5-HT2 receptors are partial agonists in this system and have a rank order of relative intrinsic efficacies corresponding to that seen in other systems possessing 5-HT2 receptors. There is an essentially linear occupancy-response relationship for 5-HT and other agonists indicating the absence of a strong amplification mechanism between receptor activation and inositol phosphate formation. The selective blockade of the 5-HT response by nanomolar concentrations of 5-HT2 selective antagonists but not by drugs acting at other 5-HT receptor subtypes suggest that the receptor in WRK1 cells is of the 5-HT2 type. Additionally, we demonstrate that in WRK1 membranes 5-HT acts via the 5-HT2 receptor to elicit a GTP dependent increase in the production of inositol-bisphosphate and inositol-trisphosphate. These properties of the WRK1 cell line indicate that it is a useful model with which to study the nature of 5-HT receptor coupling to the putative second messenger(s), the inositol phosphates.

The synthesis of new radiolabelled compounds and the evolution of the techniques designed to study the hormonal receptors allow a better understanding of their properties. Three types of vasopressin receptors have been described: the V1a receptor of liver and blood vessels, the V1b receptor of hypophysis and the V2 receptor of kidney. Such a classification was based on two criteria: The structure of the binding site and the nature of the second messenger produced. The V2 receptor coupled positively to adenylate cyclase regulates the water reabsorption via the increase of intracellular cyclic AMP. The V1a and V1b receptors involved in glycogenolysis, contraction and probably neurotransmission mobilize intracellular calcium via a positive coupling to phospholipase C. These two receptors exhibit different recognition patterns for vasopressin analogues. In mammals, the oxytocin receptors are mainly involved in myometrial contraction and lactation. Their characterization are generally difficult since they also interact with vasopressin and are sometimes colocated with vasopressin receptors. As for V1 receptor, they are coupled to phospholipase C and mobilized intracellular calcium. The receptors of angiotensin II regulate the blood pressure by different mechanisms. They are coupled to at least two transduction mechanisms (positive coupling to phospholipase C and negative coupling to adenylate cyclase). Electrophysiological data seems to indicate that such receptor may also control a calcium channel. Yet different molecules (cAMP, calcium, inositol phosphates, diacyl-glycerol) trigger the hormonal effect of angiotensin II inside the cell.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenylate cyclase has been found in membranes from all animal cells studied thus far. The catalytic activity of the enzyme can be modified in vitro by a large variety of chemical and physical agents. Some of these agents are active on adenylate cyclases from almost all cell types; examples of such agents are sodium fluoride (1) and to a lesser extent guanylnucleotides (2). Other agents, e. g., many peptide hormones, several neurotransmitters, and prostaglandins are active on the adenylate cyclase from one or a limited number of cell types. It is generally accepted that the specificity of the response is due to the presence on the membrane of specific receptor molecules. These receptors are able (1) to bind the regulatory agent and (2) to initiate the sequence of molecular events leading to a change (in most cases an increase) in the catalytic activity of adenylate cyclase. In the past few years considerable effort has been devoted to the characterization of membrane receptors functionally coupled to adenylate cyclase (for review, see Ref. 3). Some of these receptors have been extracted from the membrane in an active form. However, in none of the receptor adenylate cyclase systems investigated so far were the receptors present in a sufficiently great amount to allow complete purification and physicochemical characterization. On the other hand, very little is known about adenylate cyclase itself and about the molecular mechanisms involved in its activation by specific and nonspecific regulatory agents Almost all available data were derived from kinetic studies using intact membranes. Adenylate cyclases from various sources were found to be extractable from the membrane under the influence of nonionic detergents (4–6). However, it was not possible to obtain a purified and stable form of the soluble enzyme. Furthermore, the receptor-mediated enzyme activation was lost during membrane solubilization.

WRK1 cells, an established cell line derived from a chemically induced mammary tumor in the rat, are sensitive to vasopressin. Binding studies with intact WRK1 cells indicated the presence of a single population of [3H]vasopressin binding sites (dissociation constant, Kd = 12.7 +/- 0.2 nM, maximal binding capacity = 75 +/- 6 fmole/10(6) cells). Competition experiments using a series of vasopressin analogs with enhanced selectivity for the three subtypes of receptors already characterized--that is, renal V2 receptors, V1 receptors of the vascular or hepatic subtype (V1a), and V1 receptors from rat adenohypophysis (V1b)--indicated that vasopressin receptors from WRK1 cells have a ligand specificity very similar, if not identical, to that of V1a receptors. Vasopressin induced a marked (up to tenfold) increase in the production of labeled inositol phosphate (Ins 1,4,5 P3, Ins 1,4 P2, and Ins P) by WRK1 cells prelabeled with [3H]inositol. Antagonists of the vasopressor effect of vasopressin inhibited vasopressin-induced inositol lipid breakdown in WRK1 cells. For the entire series of vasopressin analogs tested, there was a close correlation between the respective Kd values for binding of these peptides to WRK1 cells and the corresponding Ka or Ki values derived from the determination of dose-dependent stimulation of inositol phosphate production, or inhibition of vasopressin-induced stimulation.

Hormones, neurotransmitters and other signaling molecules that mobilize intracellular calcium stimulate the activity of the phospholipase C. This enzyme specifically hydrolyses the phosphoinositides and leads to the generation of two second messengers: the inositol triphosphate which mobilizes calcium from the endoplasmic pools and the diacylglycerol which stimulates the protein kinase C. The integration of these two metabolic pathways lead to the biological effect associated to the hormone or the neurotransmitter considered. In this review we will focussed our attention on the first step of these transducing mechanisms: the receptor-mediated hydrolysis of phosphoinositides.

Despite their opposite effects on prolactin secretion, both dopamine and angiotensin II inhibit adenylate cyclase activity in homogenates of anterior pituitary cells in primary culture. Dopamine and angiotensin II inhibition of adenylate cyclase was not additive, suggesting that both neurohormones inhibit the adenylate cyclase of the lactotroph cells. Pretreatment with Bordetella pertussis toxin (islet activator protein) completely suppressed the dopamine-induced inhibition of both adenylate cyclase and prolactin secretion. The islet activator protein also reversed the angiotensin II-induced inhibition of the adenylate cyclase activity. In contrast, angiotensin II stimulation of prolactin release was not affected by the toxin. Angiotensin II also induced a dose-dependent stimulation of inositol phosphates (250%) with an EC50 of 0.1 nM, close to that observed for prolactin secretion. Islet activator protein pretreatment did not block the stimulation of inositol phosphate production. Dopamine inhibited the angiotensin II-stimulated prolactin release and the production of inositol phosphates induced by angiotensin II. It is concluded that angiotensin II and dopamine receptors of lactotroph cells are able to modulate both cAMP and inositol phosphate production. The dopamine receptor of lactotrophs appears to be the first example of a receptor which is negatively coupled to the production of inositol phosphates.

The use of mesterolone (30-75 mg daily for 60 days) in the treatment of primary and secondary male sterility (oligospermia, asthenospermia, a bnormal sperm viscosity, limited sperm survival time) in 141 patients is reported. Improvement of at least 1 symptom was seen in 44 patients. In 19 cases, the spouse became pregnant. The lower dose was associated with a somewhat higher improvement rate. Tolerance of mesterolone was excellent; side effects were observed in 2 patients. It is concluded that mesterolone is a useful addition to the therapeutic arsenal for the treatment of male infertility.

The effects of congenital hypothyroidism on both the structure and function of the renal medulla were studied by comparing, in 1-month old rats, the structural features of collecting tubules with the capacity of vasopressin to bind membrane preparations and the related adenylate cyclase activation. With the exception of a reduced caliber, hypothyroidism had no effect on the density, total number, distribution of tubules according to epithelial thickness, or on the number of epithelial cells, or their area. The binding capacity of vasopressin and the related adenylate cyclase activation were equally reduced by about 50%, without changes in (i) the basal or guanylyl-imidodiphosphate (Gpp(NH)p)-stimulated adenylate cyclase activities, (ii) the apparent dissociation constant (KD) of labelled vasopressin from its specific receptor or (iii) the apparent activation constant (Kact) of vasopressin for adenylate cyclase. Taken together, these results clearly demonstrate that congenital hypothyroidism exerts a direct influence on the developing responsiveness of the renal medulla, mainly by reducing the density of active hormone receptors per cell, instead of reducing cell number or cell membrane area.

Transducin (T), the GTP-binding protein of the retina activates the cGMP phosphodiesterase system, and presents analogies with the proteins GS and Gi which respectively mediate adenylate cyclase activation and inhibition by hormone receptors. These proteins are all comprised of an alpha subunit carrying the GTP-binding site and a beta gamma subunit made of two peptides. The beta peptide (35 kd) appears similar in the three proteins. We demonstrate here that purified T beta gamma inhibits adenylate cyclase from human platelet membranes. This inhibition was observed when adenylate cyclase was stimulated by GTP, prostaglandin E1 (PGE1), NaF and forskolin, but not when stimulated by GTP(gamma)S. In the presence of GTP and forskolin, the T beta gamma-induced maximal inhibition was not additive with the alpha 2-receptor-induced adenylate cyclase inhibition mediated by Gi. Both inhibitions were suppressed at high Mg2+ concentrations, which as also known to dissociate T beta gamma from T alpha-GDP. This suggests that these adenylate cyclase inhibitions are due to the formation of inactive complexes of GS alpha-GDP with T beta gamma or Gi beta gamma. T beta gamma-induced inhibition did not require detergent and could be suppressed by simple washing. T beta gamma effects are dependent on its concentration rather than on its total amount. This suggests that T beta gamma can operate in solution with no integration into the membrane. Similar inhibitory effects of T beta gamma are observed on adenylate cyclase from anterior pituitary and lymphoma S49 cell lines.

The angiotensin II analogues, [Sar1, (4'-N3)Phe8]All and [Sar1, (4'-N3)D-Phe8]All were synthetized and prepared in their iodinated and radioiodinated forms. On rat liver membranes 125I-SarN3PheAll and 125I-SarN3DPheAll labeled a single population of sites (maximal binding capacity, 1.13 +/- 0.16 pmol/mg of protein) with high affinity (dissociation constants of 0.18 +/- 0.05 and 0.37 +/- 0.16 nM, respectively) and high specificity (nonspecific binding less than 10% of total binding for an 0.8 fractional receptor occupancy). 125I-SarN3PheAll and 125I-SarN3DPheAll allowed photoaffinity labeling of liver angiotensin receptors with high efficiency. The yield of photoaffinity labeling was independent of fractional receptor occupancy. 125I-SarN3PheAll and 125I-SarN3DPHeAll allow recovery in a soluble and covalently labeled form of about 40% of the total number of angiotensin receptors from rat liver membranes. For this reason they can be considered as potentially very useful tools for the purification of angiotensin receptors. The material covalently labeled with the antagonist 125I-SarN3DPHeAll was eluted from an Ultrogel ACA34 column as a homogeneous peak (Stoke's radius: 5.5 +/- 0.2 nm). The material covalently labeled with the agonist 125I-SarN3PHeAll was more heterogeneous. Sodium dodecylsulfate-polyacrylamide gel electrophoresis of 125I-SarN3PheAll- and 125I-SarN3DPHeAll-labeled material revealed a single component of 63,000 molecular weight.

By means of scanning electron-microscopic investigations, thorn-like projections (TLP) were observed in the cilio-secretory epithelium of the ventral surface of the human endocervix. These projections, which seem to be characteristic of a new cell type, were seen at different stages of the menstrual cycle (days 8, 14 and 21) in 4 apparently healthy fertile women. Striking differences in length, with a maximum size at midcycle, suggest an evolutive process throughout the menstrual cycle. The origin and possible physiological role of TLP in the reproductive process are discussed.

The loading and loss kinetics of cellular 45Ca were studied in rat liver cells. The data are described by a three compartment system. Six arrangements of the exchange pathways of these pools are presented. Taking into account the Ca stocking properties of cell organels, two models may be emphasized.