Role of the Δ Resonance in the Population of a Four-Nucleon State in the

The ^{54}Fe nucleus was populated from a ^{56}Fe beam impinging on a Be target with an energy of E/A=500 MeV. The internal decay via γ-ray emission of the 10^{+} metastable state was observed. As the structure of this isomeric state has to involve at least four unpaired nucleons, it cannot be populated in a simple two-neutron removal reaction from the ^{56}Fe ground state. The isomeric state was produced in the low-momentum (-energy) tail of the parallel momentum (energy) distribution of ^{54}Fe, suggesting that it was populated via the decay of the Δ^{0} resonance into a proton. This process allows the population of four-nucleon states, such as the observed isomer. Therefore, it is concluded that the observation of this 10^{+} metastable state in ^{54}Fe is a consequence of the quark structure of the nucleons.

Isospin Mixing in

The isospin mixing was deduced in the compound nucleus ^{80}Zr at an excitation energy of E^{*}=54 MeV from the γ decay of the giant dipole resonance. The reaction ^{40}Ca+^{40}Ca at E_{beam}=136 MeV was used to form the compound nucleus in the isospin I=0 channel, while the reaction ^{37}Cl+^{44}Ca at E_{beam}=95 MeV was used as the reference reaction. The γ rays were detected with the AGATA demonstrator array coupled with LaBr_{3}:Ce detectors. The temperature dependence of the isospin mixing was obtained and the zero-temperature value deduced. The isospin-symmetry-breaking correction δ_{C} used for the Fermi superallowed transitions was extracted and found to be consistent with ß-decay data.

Isospin character of low-lying pygmy dipole states in 208Pb via inelastic scattering of 17O ions.

The properties of pygmy dipole states in 208Pb were investigated using the 208Pb(17O, 17O'γ) reaction at 340 MeV and measuring the γ decay with high resolution with the AGATA demonstrator array. Cross sections and angular distributions of the emitted γ rays and of the scattered particles were measured. The results are compared with (γ, γ') and (p, p') data. The data analysis with the distorted wave Born approximation approach gives a good description of the elastic scattering and of the inelastic excitation of the 2+ and 3- states. For the dipole transitions a form factor obtained by folding a microscopically calculated transition density was used for the first time. This has allowed us to extract the isoscalar component of the 1- excited states from 4 to 8 MeV.

Structural evolution in the neutron-rich nuclei ¹°6Zr and ¹°8Zr.

The low-lying states in ¹°6Zr and ¹°8Zr have been investigated by means of ß-γ and isomer spectroscopy at the radioactive isotope beam factory (RIBF), respectively. A new isomer with a half-life of 620 ± 150 ns has been identified in ¹°8Zr. For the sequence of even-even Zr isotopes, the excitation energies of the first 2⁺ states reach a minimum at N = 64 and gradually increase as the neutron number increases up to N = 68, suggesting a deformed subshell closure at N = 64. The deformed ground state of ¹°8Zr indicates that a spherical subshell gap predicted at N = 70 is not large enough to change the ground state of ¹°8Zr to the spherical shape. The possibility of a tetrahedral shape isomer in ¹°8Zr is also discussed.

ß-decay half-lives of very neutron-rich Kr to Tc isotopes on the boundary of the r-process path: an indication of fast r-matter flow.

The ß-decay half-lives of 38 neutron-rich isotopes from (36)Kr to (43)Tc have been measured; the half-lives of (100)Kr, (103-105)Sr, (106-108)Y, (108-110)Zr, (111,112)Nb, (112-115)Mo, and (116,117)Tc are reported here. The results when compared with previous standard models indicate an overestimation in the predicted half-lives by a factor of 2 or more in the A≈110 region. A revised model based on the second generation gross theory of ß decay better predicts the measured half-lives and suggests a more rapid flow of the rapid neutron-capture process (r-matter flow) through this region than previously predicted.

Search for the pygmy dipole resonance in 68Ni at 600 MeV/nucleon.

The gamma decay from Coulomb excitation of 68Ni at 600 MeV/nucleon on a Au target was measured using the RISING setup at the fragment separator of GSI. The 68Ni beam was produced by a fragmentation reaction of 86Kr at 900 MeV/nucleon on a 9Be target and selected by the fragment separator. The gamma rays produced at the Au target were measured with HPGe detectors at forward angles and with BaF2 scintillators at backward angles. The measured spectra show a peak centered at approximately 11 MeV, whose intensity can be explained in terms of an enhanced strength of the dipole response function (pygmy resonance). Such pygmy structure has been predicted in this unstable neutron-rich nucleus by theory.

Probing the order-to-chaos region in superdeformed 151Tb and 196Pb nuclei with continuum gamma transitions.

The gamma decay associated with the warm rotation of the superdeformed nuclei 151Tb and 196Pb has been measured with the EUROBALL IV array. Several independent quantities provide a stringent test of the population and decay dynamics in the superdeformed well. A Monte Carlo simulation of the gamma decay based on microscopic calculations gives remarkable agreement with the data only assuming a large enhancement of the B(E1) strength for 1-2 MeV gamma rays, which may be related to the evidence for octupole vibrations in both mass regions.

Giant dipole resonance in the hot and thermalized 132Ce nucleus: damping of collective modes at finite temperature.

The gamma decay of the giant dipole resonance (GDR) in the 132Ce compound nucleus with temperature up to approximately 4 MeV has been measured, using the reaction 64Ni + 68Zn at E(beam) = 300, 400, and 500 MeV. The gamma and charged particles measured in coincidence with recoils are consistent with a fully equilibrated compound nucleus emission. The GDR width, obtained with the statistical model analysis, is found to increase almost linearly with temperature. This increase is rather well reproduced within a model including thermal shape fluctuations and the lifetime of the compound nucleus.

Compound and rotational damping in warm deformed rare-earth nuclei.

The gamma decay in the quasicontinuum from selected configurations of the rotational nucleus 163Er has been measured with the EUROBALL array. A new analysis technique has allowed for the first time to directly measure the compound and rotational damping widths Gamma (micro) and Gamma (rot). Values of Gamma (micro) approximately 20 keV and Gamma (rot) approximately 200 keV are obtained in the spin region I approximately 30-40 variant Planck's over 2pi, in good agreement with microscopic cranked shell model calculations. A dependence of Gamma (micro) and Gamma (rot) on the K-quantum number of the nuclear states is also presented.

A small G-protein involved in phosphatidylinositol-4-phosphate kinase activation.

A guanine nucleotide binding protein tentatively designated Gs(PIPK) which activates purified phosphatidylinositol-4-phosphate kinase in vitro, has been partially purified from rat liver membranes and identified as a small G-protein with a molecular mass between 20 and 25 kDa.

Purification and partial characterization of phosphatidylinositol-4-phosphate kinase from rat liver plasma membranes. Further evidence for a stimulatory G-protein.

Phosphatidylinositol 4-phosphate (PIP) kinase (E.C. 2.7.1.68) has been purified about 1200-fold from rat liver plasma membranes, taking advantage of affinity chromatography on quercetin-Sepharose as a novel step. The purified PIP kinase showed no contamination by the following enzyme activities: phosphatidylinositol (PI) kinase (EC 2.7.1.67), protein kinase C (EC 2.7.1.-), diacylglycerol kinase (EC 2.7.1.-), phospholipase C (EC 3.1.4.11), protein-tyrosine kinase (EC 2.7.1.112), alkaline phosphatase (EC 3.1.3.1), triphosphoinositide phosphomonoesterase (EC 3.1.3.36), adenylate kinase (EC 2.7.4.3) and cAMP-dependent protein kinase (EC 2.7.1.37). The liver membrane enzyme requires high Mg2+ concentrations with a KM value of 10 mM. Ca2+ or Mn2+ could replace Mg2+ to a certain, though small, extent. Apparent KM values with respect to PIP and ATP were 10 and 65 microM, respectively. GTP was slightly utilized by the kinase as phosphate donor while CTP was not. Quercetin inhibited the enzyme with Ki = 34 microM. Extending our previous observations (Urumow, T. and Wieland, O.H. (1986) FEBS Lett. 207, 253-257 and Urumow, T. and Wieland, O.H. (1988) Biochim. Biophys. Acta 972, 232-238) [gamma S]pppG still stimulated the PIP kinase in extracts of solubilized liver membranes. 20-40% (NH4)2SO4 precipitation of the membrane extracts yielded a fraction that contained the bulk of enzyme activity but did not respond to stimulation by [gamma S]pppG any longer. This was restored by recombination with a protein fraction collected at 40-70% (NH4)2SO4 saturation, presumably containing a GTP binding protein and/or some other factor separated from the PIP kinase. In the reconstituted system [gamma S]pppG stimulated PIP kinase in a concentration dependent manner with maximal activation at 5 microM. This effect was not mimicked by [gamma S]pppA and was blocked by [beta S]ppG. These results strongly support our view that in liver membranes PIP kinase is regulated by a G-protein.

Protein glycation: measurement and clinical relevance.

This review of protein glycation deals with the biochemical background of glycated blood proteins, their methods of determination and their clinical significance. General reaction principles for determination of glycated proteins are discussed with special emphasis on the determination of glycated serum proteins in the clinical laboratory. Binding methods like boronate affinity, immunoassay, phenylhydrazine binding and ion exchange chromatography leave the analyte intact, whereas chemical methods like strong or mild hydrolysis or reduction in alkaline medium (fructosamine assay) results in destruction of the glycated protein. As most reactions are nonstoichiometric (except ion exchange chromatography and periodate oxidation), varying results are obtained from laboratory to laboratory. Up to now boronate affinity chromatography, mild hydrolysis yielding hydroxymethylfurfural and the fructosamine assay have been mostly used for determination of glycated serum proteins. The fructosamine assay appears to be most practical, because it is quick, economic and precise, but it suffers from unspecific side reactions. Although other methods like immunoassays or boronate ester formation in solution appear promising, there is currently no commercially available assay for the economic, precise and accurate determination of glycated serum protein. The clinical relevance of glycated serum protein determination is difficult to evaluate because the assays are based on different reaction principles and hence yield variable results. Nevertheless, the following conclusion may be drawn from the reports now available. i) The possibility that glycated serum proteins may discriminate better than glycated haemoglobin between "normal" and "diabetic" is still controversial. ii) Glycated serum proteins are formed faster than glycated haemoglobin, reflecting the changes in glycaemia for shorter periods of time (medium-term control). iii) It has not been yet established, using large cohorts, whether the glycated serum proteins allow the detection or exclusion of diabetes. iv) Determination of glycated serum proteins should not be considered as a substitute for the determination of glycated haemoglobin, but rather as a complementary determination, leading to the improved laboratory control of the diabetic patient.

Evidence for a cholera-toxin-sensitive G-protein involved in the regulation of phosphatidylinositol 4-phosphate kinase of rat liver membranes.

Studies on the phosphorylation of inositol phospholipids of rat liver membranes have shown that [gamma S]pppG stimulates 32P incorporation from [gamma-32P]ATP into PI and PIP. This effect appeared specific for stable GTP analogues and could not be reproduced by other compounds. ADP-ribosylation of the membranes with cholera toxin resulted in a large decrease of PIP2 without changes in the level of PIP. Since an activation of phospholipase C can be ruled out, the lowering of PIP2 is explained on the basis of an inhibition of PIP kinase (EC 2.7.1.68). From these results it appears that a novel cholera-toxin-sensitive G-protein is involved in the regulation of PIP kinase.

Separation of the protein-tyrosine kinase and phosphatidylinositol kinase activities of the human placental insulin receptor.

On immunoprecipitation using a specific antiphosphotyrosine antibody, phosphatidylinositol kinase (EC 2.7.1.67) activity was separated from the protein-tyrosine kinase (EC 2.7.1.112) activity of the wheat germ agglutinin (WGA) -purified insulin receptor from human placenta. This clearly indicates that protein-tyrosine kinase and phosphatidylinositol kinase activity do not reside on the same polypeptide chain as previously has been suggested. Quantitatively, the fraction of phosphatidylinositol kinase that was bound to WGA sepharose and eluted together with the insulin receptor amounted to 2% of the Triton X-100 soluble phosphatidylinositol kinase. The apparent Km values of the bound and unbound phosphatidylinositol kinase with respect to PI and ATP were very similar (0.4 and 0.3 mmol/l and 8 and 7 mumol/l, respectively) suggesting that the WGA-bound phosphatidylinositol kinase is not a different enzyme, but rather represents a small portion of the bulk Triton X-100-soluble phosphatidylinositol kinase that is bound to the lectin tightly associated with the insulin receptor. The synthetic polymer (Glu80Tyr20)n, a model substrate of the insulin receptor tyrosine kinase, at 0.5 mmol/l, inhibited phosphatidylinositol kinase of WGA-purified insulin receptor by 70-90%. This inhibition was not overcome by increasing the concentrations of ATP or PI as one would expect if a functional interrelationship of the protein-tyrosine kinase and the phosphatidylinositol kinase would exist.

Enzymatic assay of myo-inositol in serum.

An enzymatic spectrophotometric method for determination of myo-inositol in serum is described. The method is sensitive and rapid to perform without special equipment. Linearity between the amount of myo-inositol and absorbance was obtained in the range of 0.5 to 3 nmol myo-inositol per assay. The amounts of myo-inositol determined in sera from apparently healthy subjects agree well with gas chromatographic data.

An Mr 180,000 protein is an endogenous substrate for the insulin-receptor-associated tyrosine kinase in human placenta.

The beta-subunit of the insulin receptor contains a tyrosine-specific protein kinase. Insulin binding activates this kinase and causes phosphorylation of the beta-subunit of the insulin receptor. It is believed that phosphorylation of other proteins might transmit the insulin signal from the receptor to the cell. In the present study we used a polyclonal anti-phosphotyrosine antibody to detect other proteins that become tyrosine phosphorylated upon insulin stimulation. Glycoproteins from human placenta membranes were enriched by wheat germ agglutinin chromatography and phosphorylation was studied with [gamma-32P]ATP and insulin in vitro. Phosphorylated proteins were immunoprecipitated by antibodies against the insulin receptor and by serum containing the anti-phosphotyrosine antibody. Beside the insulin-stimulated phosphorylation of the 95 kDa beta-subunit of the insulin receptor, an insulin-stimulated phosphorylation of a 180 kDa protein was found. The phosphorylation of both proteins occurred only on tyrosine residues. Insulin increased 32P incorporation into the 180 kDa band 2.7-fold (S.E.M. +/- 0.3, n = 5). The 180 kDa protein was not precipitated by antibodies against the insulin receptor. H.p.l.c. chromatograms of tryptic fragments of the phosphorylated 180 kDa protein and of the beta-subunit of the insulin receptor revealed different patterns for both proteins. Insulin-stimulated phosphorylation of the 180 kDa protein was also detectable in unfractionated detergent-solubilized membranes. The phosphorylation of the 180 kDa protein was stimulated by insulin with the same dose-response curve as the phosphorylation of the beta-subunit, suggesting that this protein might be another endogenous substrate of the insulin receptor kinase.

Kinetic analysis of glycation as a tool for assessing the half-life of proteins.

Glycation of proteins, a common postribosomal modification, proceeds via Amadori rearrangement to yield a stable ketoamine linkage of glucose with the protein. Kinetic analysis of the reaction shows that the amount of glycation at steady state is proportional to the glucose concentration, to protein half-life and to the rate of glycation. Thus, when the rate of glycation is determined in vitro and the extent of glycation of a given protein isolated from euglycemic subjects is measured, the half-life may be calculated. As the in vivo situation may not be simulated accurately in vitro, the calculated values may be considered as approximation. When the calculated values were compared with values reported in the literature fairly good agreement was found except for hemoglobin. Studies on stability of glycated albumin show that ketoamine decreases by about 20% when incubated under physiological conditions for 20 days. The method described by us is especially valuable when turnover of proteins in normal and pathophysiological states are compared. The half-life of plasma low-density lipoprotein is longer in patients with hypothyroidism or a high plasma low-density lipoprotein level than in normal subjects. Extending our studies to tissue proteins we did not find a significant increase in half-life of tendon collagen with age. Basement membrane collagen turnover is faster in diabetic patients in bad metabolic control. Thus, the procedure using fructosylamine as endogenous label of protein offers a method of great potential to study the turnover of human body proteins.

Stimulation of phosphatidylinositol 4-phosphate phosphorylation in human placenta membranes by GTP gamma S.

In human placenta membranes the rate limiting enzyme for PIP2 formation from PI is PIP kinase. GTP gamma S is shown to activate PIP kinase by increasing Vmax of the enzyme. It is suggested that a guanine nucleotide regulatory protein is involved in the activation of PIP kinase although coupling with a specific receptor is not yet known. Since PIP2 is the preferred substrate of phospholipase C, the possibility exists that an increase of PIP2 due to activation of PIP kinase leads to an enhancement of phospholipase C activity and hence to an increased production of IP3 and DAG.