Biomedical subjects

M Hirasawa

Publications and source records attributed to M Hirasawa.

At least 19 recordsLinked to original sources

PubMed2004

Effect of compensation for scattering angular uncertainty in analytical Compton camera reconstruction.

In Compton cameras, the measured scattering angle is associated with an uncertainty which becomes larger as the incident gamma-ray energy decreases. Since this uncertainty degrades the spatial resolution of reconstructed images, Hirasawa and Tomitani (2003 Phys. Med. Biol. 48 1009-26) previously revised their analytical reconstruction algorithm to compensate for it. As the new algorithm improved the spatial resolution in effect, they expected an enhancement of the statistical noise. In this paper, the effect of this compensation has been analysed in view of spatial resolution (the FWHM of the noise-free reconstructed image for a point source distribution), statistical noise (the relative standard deviation of reconstructed images for an isotropic source distribution) and image quality (the roughness of reconstructed images for a phantom). The results describe not only the effect of the compensation, but also the relation between the statistical noise and three parameters, i.e., the incident gamma-ray energy, the spatial resolution and the measured total event numbers, in reconstruction with compensation. This relation should be taken into account for the design of Compton cameras with good quality images, i.e., useful image, output.

Algorithms↗

PubMed2003

An analytical image reconstruction algorithm to compensate for scattering angle broadening in Compton cameras.

Compton cameras have been developed for use in gamma-ray astronomy and nuclear medicine. Their defining merit is that they do not need collimators; however, on the demerit side, they need inversion procedures for image reconstruction, since a measured datum is proportional to the integration of incident gamma rays along a cone surface with the same Compton scattering angle. First, an iteration method was adopted for this task. Later, analytical methods were found under restricted conditions. Parra (2000 IEEE Trans. Nucl. Sci. 47 1543-50) deduced a purely analytical reconstruction algorithm for a complete set of scattering-projection data that include data at all the scattering angles. Tomitani and Hirasawa (2002 Phys. Med. Biol. 47 2129-45) found that by making a slight modification, Parra's algorithm could be extended to the scattering-projection data in limited scattering angles. However, their algorithm neglected the effects of practical problems that cause the degradation of spatial resolution. Sources of degradation were identified as noise in the energy signal of their front detector and the Doppler effect in the scattering process. In this paper, we first analyse the effects of these sources on the angular resolution of the scattering-projection data and then present a revised reconstruction algorithm in which these two factors are incorporated. Simulation studies on digital phantoms reveal that the algorithm can reconstruct images even when these two factors are included.

Algorithms↗

PubMed2002

Image reconstruction from limited angle Compton camera data.

The Compton camera is used for imaging the distributions of gamma ray direction in a gamma ray telescope for astrophysics and for imaging radioisotope distributions in nuclear medicine without the need for collimators. The integration of gamma rays on a cone is measured with the camera, so that some sort of inversion method is needed. Parra found an analytical inversion algorithm based on spherical harmonics expansion of projection data. His algorithm is applicable to the full set of projection data. In this paper, six possible reconstruction algorithms that allow image reconstruction from projections with a finite range of scattering angles are investigated. Four algorithms have instability problems and two others are practical. However, the variance of the reconstructed image diverges in these two cases, so that window functions are introduced with which the variance becomes finite at a cost of spatial resolution. These two algorithms are compared in terms of variance. The algorithm based on the inversion of the summed back-projection is superior to the algorithm based on the inversion of the summed projection.

Algorithms↗

PubMed2002

Modulation of synaptic transmission by oxytocin and vasopressin in the supraoptic nucleus.

It is now generally accepted that magnocellular neurons of the supraoptic and paraventricular nuclei release the neuropeptides oxytocin and vasopressin from their dendrites. Peptide release from their axon terminals in the posterior pituitary and dendrites differ in dynamics suggesting that they may be independently regulated. The dendritic release of peptide within the supraoptic nucleus (SON) is an important part of its physiological function since the local peptides can regulate the electrical activity of magnocellular neurons (MCNs) which possess receptors for these peptides. This direct postsynaptic action would affect the output of peptide in the neurohypophysis. Another way that these peptides can regulate MCN activity would be to modulate afferent inputs unto themselves. Although the influence of afferent inputs (inhibitory and excitatory) on SON magnocellular neuron physiology has been extensively described in the last decade, a role for these locally released peptides on synaptic physiology of this nucleus has been difficult to show until recently, partly because of the difficulty of performing stable synaptic recordings from these cells in suitable preparations that permit extensive examination. We recently showed that under appropriate conditions, oxytocin acts as a retrograde transmitter in the SON. Oxytocin, released from the dendrites of MCNs, decreased evoked excitatory synaptic transmission by inhibiting glutamate release from the presynaptic terminals. It modulated voltage-dependent calcium channels, mainly N-type and to a lesser extent P/Q-type channels, located on glutamatergic terminals. Although evidence is less conclusive, it is possible that vasopressin has similar actions to reduce excitatory transmission. This synaptic depressant effect of oxytocin and/or vasopressin, released from dendrites, would ensure that MCNs regulate afferent input unto themselves using their own firing rate as a gauge. Alternatively, it may only be a subset of afferent terminals that are sensitive to these peptides, thereby providing a means for the MCNs to selectively filter their afferent inputs. Indeed its specificity is partly proven by our observation that oxytocin does not affect spontaneous glutamate release, or GABA release from inhibitory terminals (Brussaard et al., 1996). Thus, the dendrites of MCNs of the supraoptic nucleus serve a dual role as both recipients of afferent input and regulators of the magnitude of afferent input, allowing them to directly participate in the shaping of their output. This adds to a rapidly growing body of evidence in support of the concept of a two-way communication between presynaptic terminals and postsynaptic dendrites, and shows the potential of this nucleus as a model to study such form of synaptic transmission.

Animals↗

PubMed2002

[Surgery for thymomas and thymic carcinomas; treatment results in terms of WHO histologic typing, Masaoka staging system, and p53 expression].

Forty thymomas and thymic carcinomas were classified in terms of WHO histologic typing, Masaoka staging system, and p53 expression. In WHO histologic typing, type A, AB, B1, B2, B3, and C were 1, 10, 16, 5, 4, and 4 cases, respectively. In Masaoka staging system, I, II, III, and IV were 15, 9, 10, and 6 cases, respectively. Thirteen thymomas exhibited positive p53 expression and 27 did not. Type A and AB thymomas had more favorite prognosis than type B3 and C thymomas, and prognosis of type B1 and B2 was middle. Staging by the Masaoka system also correlated with survival rates. Patients who had p53-negative thymomas survived longer than those who had p53-positive thymomas. A treatment strategy for thymomas and thymic carcinomas should be made on the basis of WHO histologic typing, Masaoka staging system, and p53 expression.

Adult↗

PubMed2002

Lipo-prostaglandin E1 is absorbed by vascular smooth muscle cells and may enhance re-endothelialization of vein grafts.

BACKGROUND: Enhancement of re-endothelialization inhibits the progression of intimal hyperplasia. We investigated uptake of Lipoprostaglandin E(1) (LPGE(1)) by endothelial cells (ECs) and vascular smooth muscle cells (VSMCs), and effects of the LPGE(1) on re-endothelialization of vein grafts. METHODS: Primary cultured ECs and VSMCs were obtained from mongrel dogs, and incubated in mediums containing 0.5, 5, 50, and 250 ng/ml of DiI-LPGE(1) (DLPGE(1)); its uptake was visualized by the standard rhodamine excitation, and assessed by flow cytometry. In an implantation study, the bilateral femoral veins of 18 animals were implanted into the femoral artery, and the animals were given 0.2 microg/kg LPGE(1) for 2 weeks. Percentages of EC coverage area of the grafts (%EC) were measured by silver nitrate staining at intervals of 3, 7, and 21 days after implantation. RESULTS: VSMCs showed significant uptake in all of the mediums containing DLPGE(1), whereas the ECs revealed no fluorescence. In flow cytometry, histograms of VSMCs showed a specific notch of DLPGE(1), which increased the height according to the grade of the concentrations. The notch did not appear in the histograms of the ECs incubated with any concentration nor in the VSMCs incubated in control medium. These results suggested that LPGE(1) is predominantly absorbed by the VSMCs and exuded PGE(1) then acts on the VSMCs itself and on ECs alike. The %EC at 7 days was 58.4+/-1.9% in the DLPGE(1) group and 31.6+/-6.8% in controls (p<0.05) showing a significant enhancing effect of DLPGE(1) on re-endothelialization of the vein grafts. CONCLUSIONS: In an animal model, daily administration of LPGE(1) resulted in significant enhancement of vein graft re-endothelialization. LPGE(1) was absorbed by the VSMCs, whereas the ECs showed no uptake, therefore the enhancement is probably due to a paracrine effect of VSMCs.

Alprostadil↗

PubMed2001

Oxidation-reduction and activation properties of chloroplast fructose 1,6-bisphosphatase with mutated regulatory site.

The concentration of Mg(2+) required for optimal activity of chloroplast fructose 1,6-bisphosphatase (FBPase) decreases when a disulfide, located on a flexible loop containing three conserved cysteines, is reduced by the ferredoxin/thioredoxin system. Mutation of either one of two regulatory cysteines in this loop (Cys155 and Cys174 in spinach FBPase) produces an enzyme with a S(0.5) for Mg(2+) (0.6 mM) identical to that observed for the reduced WT enzyme and significantly lower than the S(0.5) of 12.2 mM of oxidized WT enzyme. E(m) for the regulatory disulfide in WT spinach FBPase is -305 mV at pH 7.0, with an E(m) vs pH dependence of -59 mV/pH unit, from pH 5.5 to 8.5. Aerobic storage of the C174S mutant produces a nonphysiological Cys155/Cys179 disulfide, rendering the enzyme partially dependent on activation by thioredoxin. Circular dichroism spectra and thiol titrations provide supporting evidence for the formation of nonphysiological disulfide bonds. Mutation of Cys179, the third conserved cysteine, produces FBPase that behaves very much like WT enzyme but which is more rapidly activated by thioredoxin f, perhaps because the E(m) of the regulatory disulfide in the mutant has been increased to -290 mV (isopotential with thioredoxin f). Structural changes in the regulatory loop lower S(0.5) for Mg(2+) to 3.2 mM for the oxidized C179S mutant. These results indicate that opening the regulatory disulfide bridge, either through reduction or mutation, produces structural changes that greatly decrease S(0.5) for Mg(2+) and that only two of the conserved cysteines play a physiological role in regulation of FBPase.

Catalytic Domain↗

PubMed2001

Binding features of chloroplast fructose-1,6-bisphosphatase-thioredoxin interaction.

It has been proposed that a hydrophobic groove surrounded by positively charged amino acids on thioredoxin (Trx) serves as the recognition and docking site for the interaction of Trx with target proteins. This model for Trx-protein interactions fits well with the Trx-mediated fructose-1,6-bisphosphatase (FBPase) activation, where a protruding negatively charged loop of FBPase would bind to this Trx groove, in a process involving both electrostatic and hydrophobic interactions. This model facilitates the prediction of Trx amino acid residues likely to be involved in enzyme binding. Site-directed mutagenesis of some of these amino acids, in conjunction with measurements of the FBPase activation capacity of the wild type and mutated Trxs, was used to check the model and provided evidence that lysine-70 and arginine-74 of pea Trx m play an essential role in FBPase binding. The binding parameters for the interaction between chloroplast FBPase and the wild type pea Trxs f and m, as well as mutated pea Trx m, determined by equilibrium dialysis in accordance with the Koshland-Nemethy-Filmer model of saturation kinetics, provided additional support for the role of these basic Trx residues in the interaction with FBPase. These data, in conjunction with the midpoint redox potential (E(m)) determinations of Trxs, support the hydrophobic groove model for the interaction between chloroplast FBPase and Trx. This model predicts that differences in the FBPase activation capacity of Trxs arise from their different binding abilities.

Arginine↗

PubMed2001

Oxytocin retrogradely inhibits evoked, but not miniature, EPSCs in the rat supraoptic nucleus: role of N- and P/Q-type calcium channels.

We previously reported that oxytocin (OXT), released from the dendrites of magnocellular neurons in the supraoptic nucleus (SON), acts retrogradely on presynaptic terminals to inhibit glutamatergic transmission. Here we test the hypothesis that oxytocin reduces calcium influx into the presynaptic terminal. We used nystatin perforated-patch recording in vitro to first identify the calcium channels involved in glutamatergic transmission in the SON. [omega]-Conotoxin GVIA ([omega]-CTx) and [omega]-Agatoxin TK ([omega]-Aga) both reduced evoked EPSC amplitude, while nicardipine and nickel had no effect. A combination of [omega]-CTx and [omega]-Aga completely abolished the evoked EPSCs. This depressant effect was accompanied by an increase in the paired pulse ratio with no change in the kinetics of the evoked EPSCs, AMPA currents or postsynaptic cell properties. These results suggest that presynaptic N- and P/Q-type calcium channels mediate glutamate release in the SON while L-, T- and R-type channels make little or no contribution. Oxytocin-induced reduction of the evoked EPSC was substantially occluded in the presence of [omega]-CTx but only partially in the presence of [omega]-Aga. Amastatin, an endopeptidase inhibitor that increases the level of endogenous OXT, also reduced the evoked EPSC. This amastatin effect was also occluded by [omega]-CTx and [omega]-Aga. Miniature EPSCs, which are independent of extracellular calcium, were unaffected by either [omega]-CTx or by OXT, thus further substantiating an action of both compounds on calcium channels. Therefore, dendritically released oxytocin acts mainly via a mechanism involving the N-type channel, and to a lesser extent the P/Q-type channel, to decrease excitatory transmission.

Agatoxins↗

PubMed2001

Synergistic contributions of cyclin-dependant kinase 5/p35 and Reelin/Dab1 to the positioning of cortical neurons in the developing mouse brain.

Cyclin-dependent kinase (Cdk) 5 is a unique member of the Cdk family, because Cdk5 kinase activity is detected only in the nervous tissue. Two neuron-specific activating subunits of Cdk5, p35 and p39, have been identified. Overlapping expression pattern of these isoforms in the embryonic mouse brain and the significant residual Cdk5 kinase activity in brain homogenate of the p35-/- mice indicate the redundant functions of the Cdk5 activators in vivo. Severe neuronal migration defects in p35-/-Cdk5 +/- mice further support the idea that the redundant expression of the Cdk5 activators may cause a milder phenotype in p35-/- mice compared with Cdk5-/- mice. Mutant mice lacking either Cdk5 or p35 exhibit certain similarities with Reelin/Dab1-mutant mice in the disorganization of cortical laminar structure in the brain. To elucidate the relationship between Cdk5/p35 and Reelin/Dab1 signaling, we generated mouse lines that have combined defects of these genes. The addition of heterozygosity of either Dab1 or Reelin mutation to p35-/- causes the extensive migration defects of cortical neurons in the cerebellum. In the double-null mice of p35 and either Dab1 or Reelin, additional migration defects occur in the Purkinje cells in the cerebellum and in the pyramidal neurons in the hippocampus. These additional defects in neuronal migration in mice lacking both Cdk5/p35 and Reelin/Dab1 indicate that Cdk5/p35 may contribute synergistically to the positioning of the cortical neurons in the developing mouse brain.

Adaptor Proteins, Signal Transducing↗

PubMed2001

Protective immunity to Streptococcus mutans induced by nasal vaccination with surface protein antigen and mutant cholera toxin adjuvant.

In this study, mice were immunized nasally with surface protein antigen of Streptococcus mutans serotype c (PAc) and a nontoxic A subunit mutant of cholera toxin (mCT) E112K, as a mucosal adjuvant. Immunization with PAc and mCT elicited significant PAc-specific secretory IgA in saliva and in nasal secretions. Antibody-forming cell (AFC) analysis confirmed the antibody (Ab) titers by revealing significant numbers of PAc-specific IgA AFCs in the submandibular gland and nasal passages. Furthermore, CD4(+) T cells from cervical lymph nodes exhibited significant proliferative responses when restimulated with PAc in vitro. Importantly, mice that were nasally immunized with PAc plus mCT E112K exhibited significantly reduced oral colonization by S. mutans. These results show that nasal administration of PAc and mCT E112K is potentially an effective mucosal vaccine against dental caries and reduces the colonization of S. mutans in the oral cavity.

Adjuvants, Immunologic↗

PubMed2001

GABA(B) receptors modulate short-term potentiation of spontaneous excitatory postsynaptic currents in the rat supraoptic nucleus in vitro.

High-frequency stimulation of afferents to the supraoptic nucleus (SON) results in a robust increase in the frequency and amplitude of pharmacologically isolated, tetrodotoxin-resistant, miniature excitatory postsynaptic currents (mEPSCs) lasting for 5-20 min. This increase in mEPSC frequency, termed short-term potentiation (STP), is tightly coupled to increases in action potential firing in magnocellular neurons (MCNs) suggesting a functional role for STP. gamma-Aminobutyric acid (GABA), acting selectively on GABA(B) receptors, has been shown to modulate action potential-dependent EPSCs, as well as mEPSCs in this nucleus. In this study, we examined the role of GABA in STP. Using in vitro hypothalamic slices containing the SON and the nystatin perforated-patch recording technique to record from MCNs, we tested the hypothesis that GABA modulates STP. Baclofen, a GABA(B) receptor agonist, caused a reversible decrease in the frequency of mEPSCs as well as a reduction in the magnitude and duration of STP. GABA(B) receptor antagonists blocked the baclofen-induced decrease in mEPSC frequency and reduction in STP. In addition, the antagonists by themselves increased basal mEPSC frequency while prolonging the duration of STP in most cells. By contrast, picrotoxin, a GABA(A) chloride channel blocker, had no effect on STP.These findings indicate that GABA is tonically present in the SON and its action at the GABA(B) receptor may determine the magnitude and duration of STP.

Animals↗

PubMed2001

Neuron-specific expression of Cre recombinase during the late phase of brain development.

Gene targeting to disrupt gene expression in a temporal and spatial manner in a specific tissue using Cre recombinase-mediated gene inactivation has been proven to be useful to study in vivo gene function. To delete genes specifically in neurons during the late phase of brain development, we have generated transgenic mouse lines that express Cre recombinase under the control of the murine neurofilament-H (mNF-H) gene promoter. In this study, we report that one of these mouse lines expresses Cre recombinase specifically in the neurons of the brain and spinal cord during the late stage of their development. The transgenic line displays specific excision of the loxP-flanked gene in the neurons just after embryonic day 18.5 (E.18.5), which coincides with the later phase of brain maturation including spinal cord and olfactory bulb area. This mNF-H-cre transgenic mouse line will be valuable for studying in vivo functions of neuron-specific genes, particularly, defining their precise roles in the mature nervous system using conditional gene targeting strategies.

Age Factors↗

PubMed2001

Morphological control of laser-synthesized silicon nanoparticles.

Size and crystallinity controlled silicon nanoparticles were prepared by a laser ablation, in situ annealing and mobility size-selection with a differential mobility analyzer (DMA). The shape and crystal structure of generated particles were observed by a transmission electron microscopy (TEM). Both densification of agglomerates and crystal growth of the particles were observed. The size of silicon primary particle was increased by the annealing, and the uniformity of the particle classified at 10 nm was improved as a result.

Crystallization↗

PubMed2000

Oxidation-reduction properties of two engineered redox-sensitive mutant Escherichia coli malate dehydrogenases.

Redox potentials for two inactivating intrasubunit disulfides that link helix-5 and helix-9 in mutant Escherichia coli malate dehydrogenases have been determined. The Em is -285 mV when cysteines are at positions 121 and 305 and -295 mV when the cysteines are at positions 122 and 305. Oxidation to the disulfide affects kcat but not Km values. In the single V121C and N122C mutants, the Cys in helix-5 affects the Km for oxalacetate. The pH optimum in the direction of malate formation is affected by the redox state of the enzyme. Clearly, a disulfide bond can and does form between Cys residues substituted into positions 121 or 122 in the nucleotide binding domain and 305 in the carbon substrate binding domain of this NAD-dependent malate dehydrogenase. Apparently, crosslinking the domains interferes with catalysis.

Catalysis↗

PubMed2000

Technique for enhanced rare earth separation

A process is demonstrated for the efficient separation of rare earth elements, using a combination of selective reduction and vacuum distillation of halides. The large differences in the redox chemistry of the rare earth elements and in the vapor pressures of rare earth di- and trihalides are exploited for separation. Experimental proof of concept is provided for the binary systems praseodymium-neodymium and neodymium-samarium. This process enhances the separation factor for the isolation of samarium and neodymium from their mixture by more than an order of magnitude.

Journal Article↗

PubMed2000

Oxidation-reduction properties of disulfide-containing proteins of the Rhodobacter capsulatus cytochrome c biogenesis system.

Oxidation-reduction titrations for the active-site disulfide/dithiol couples of the helX- and ccl2-encoded proteins involved in cytochrome c biogenesis in the purple non-sulfur bacterium Rhodobacter capsulatus have been carried out. The R. capsulatus HelX and Ccl2 proteins are predicted to function as part of a dithiol/disulfide cascade that reduces a disulfide on the apocytochromes c so that two cysteine thiols are available to form thioether linkages between the heme prosthetic group and the protein. Oxidation-reduction midpoint potential (E(m)) values, at pH 7.0, of -300 +/- 10 and -210 +/- 10 mV were measured for the HelX and Ccl2 (a soluble, truncated form of Ccl2) R. capsulatus proteins, respectively. Titrations of the disulfide/dithiol couple of a peptide designed to serve as a model for R. capsulatus apocytochrome c(2) have also been carried out, and an E(m) value of -170 +/- 10 mV was measured for the model peptide at pH 7.0. E(m) versus pH plots for HelX, Ccl2, and the apocytochrome c(2) model peptide were all linear over the pH range from 5.0 to 8.0, with the -59 mV/pH unit slope expected for a reaction in which two protons are taken up for each disulfide that is reduced. These results provide thermodynamic support for the proposal that HelX reduces Ccl2 and that reduced Ccl2, in turn, serves as the reductant for the production of the two thiols of the CysXxxYyyCysHis heme-binding motif of the apocytochromes.

Bacterial Proteins↗

PubMed2000

Inhibition of the thioredoxin-dependent activation of the NADP-malate dehydrogenase and cofactor specificity.

The chloroplastic NADP-malate dehydrogenase is activated by reduction of its N- and C-terminal disulfides by reduced thioredoxin. The activation is inhibited by NADP(+), the oxidized form of the cofactor. Previous studies suggested that the C-terminal disulfide was involved in this process. Recent structural data pointed toward a possible direct interaction between the C terminus of the oxidized enzyme and the cofactor. In the present study, the relationship between the cofactor specificity for catalysis and for inhibition of activation has been investigated by changing the cofactor specificity of the enzyme by substitution of selected residues of the cofactor-binding site. An NAD-specific thiol-regulated MDH was engineered. Its activation was inhibited by NAD(+) but no longer by NADP(+). These results demonstrate that the oxidized cofactor is bound at the same site as the reduced cofactor and support the idea of a direct interaction between the negatively charged C-terminal end of the enzyme and the positively charged nicotinamide ring of the cofactor, in agreement with the structural data. The structural requirements for cofactor specificity are modeled and discussed.

Amino Acid Sequence↗