Transportation of a radioactive ion beam for precise laser-trapping experiments.

Francium is the heaviest species among the alkali elements. Due to its properties, francium is said to be of advantage in measurements of tiny observations, such as atomic parity violation and electric dipole moment. Before executing experiments with francium, it must be produced artificially because it is one of the most unstable elements. We produced francium with the nuclear fusion reaction of an oxygen beam and gold target, ionized the produced francium through a thermal ionization process, and extracted the ion with electrostatic fields. However, the thermal ionization process is known to ionize not only an objective atom but also other atomic species. Therefore, a Wien filter was installed to analyze the composition of the ion beam and purify the beam. This allowed us to improve the beam purity from ∼10(-6) to ∼10(-3).

Laser frequency locking with 46 GHz offset using an electro-optic modulator for magneto-optical trapping of francium atoms.

We demonstrate frequency offset locking between two laser sources using a waveguide-type electro-optic modulator (EOM) with 10th-order sidebands for magneto-optical trapping of Fr atoms. The frequency locking error signal was successfully obtained by performing delayed self-homodyne detection of the beat signal between the repumping frequency and the 10th-order sideband component of the trapping light. Sweeping the trapping-light and repumping-light frequencies with keeping its frequency difference of 46 GHz was confirmed over 1 GHz by monitoring the Doppler absorption profile of I2. This technique enables us to search for a resonance frequency of magneto-optical trapping of Fr.

Further improvement of the upper limit on the direct 3α decay from the Hoyle state in 12C.

The direct 3α decay branch from the 02+ state at Ex=7.65 MeV in 12C, which is known as the Hoyle state, is considered to affect the triple-α reaction rate strongly and to give crucial information on its structure. We have performed a high-precision measurement of the 3α decay from this state using the 12C(12C,3α)12C reaction at E12C=110 MeV. The branching ratio of the direct 3α decay was under the detection limit in the present experiment. By comparing with Monte Carlo simulations for three decay mechanisms as the sequential decay through the ground state of ^{8}Be, the direct decay with equal energies of three α particles, and the direct decay to the phase space uniformly, we have obtained the upper limit of 0.2% on the direct 3α decay.

Radioactive ion beam transportation for the fundamental symmetry study with laser-trapped atoms.

The search for the violation of the fundamental symmetry in a radioactive atom is the promising candidate for precision tests of the standard model and its possible extensions. The subtle signal arising from the symmetry violation is enhanced in heavy atoms, such as a francium (Fr). To realize high precision measurements, a large amount of radioactive isotopes is required. The Fr is produced via a nuclear fusion reaction using a melted gold target with a (18)O primary beam at Cyclotron and Radioisotope Center, Tohoku University. The maximum extraction efficiency of the Fr ion was achieved at approximately 35%. The beam line consists of an electrostatic deflector, three electrostatic quadrupole triplets to the measurement area at 10 m away from the reaction point, and several beam diagnosis systems. We optimized parameters of the beam line.

Transcription factor FoxA (HNF3) on a nucleosome at an enhancer complex in liver chromatin.

Nucleosome-like particles and acetylated histones occur near active promoters and enhancers, and certain transcription factors can recognize their target sites on the surface of a nucleosome in vitro; yet it has been unclear whether transcription factors can occupy target sites on nucleosomes in native chromatin. We developed a method for sequential chromatin immunoprecipitation of distinct nuclear proteins that are simultaneously cross-linked to nucleosome-sized genomic DNA segments. We find that core histone H2A co-occupies, along with the FoxA (hepatocyte nuclear factor-3) transcription factor, DNA for the albumin transcriptional enhancer in native liver chromatin, where the enhancer is active. Because histone H2A on nuclear DNA is only known to exist in nucleosomes, we conclude that transcription factors can form a stable complex on nucleosomes at an active enhancer element in vivo.

FGF-2 immunoreactivity in adult rat ependyma and choroid plexus: responses to global forebrain ischemia and intraventricular FGF-2.

Fibroblast growth factor 2 (FGF-2) immunoreactivity (IR) was examined in the ependyma and choroid plexus (CP) of lateral and third ventricles in normal adult rats, as well as in response to transient forebrain ischemia (TFI) and exogenous FGF-2 delivered intraventricularly for several days by osmotic pump. Similar patterns of FGF-2 IR were seen in the CP epithelia of both lateral and third ventricles, as well as in ependymal cells of the third ventricle and along lateral sides of the lateral ventricles. Consistent staining was seen along the apical aspect of epithelial cells facing the cerebrospinal fluid (CSF). Cytoplasmic staining was seen in the absence of ischemia, and was dramatically reduced in response to TFI. FGF-2 treatment followed by TFI resulted in sustained FGF-2 IR within CP and ependymal cells, supporting the idea that these tissues are involved in synthesis and secretion of growth factors into the CSF. In contrast, along the medial sides of the lateral ventricles, adjacent to brain structures such as the hippocampus, consistent staining was seen along the basal aspect of the ependymal cells. We propose that at least some regions of ependyma may function to transport molecules such as FGF-2 directly into the underlying brain parenchyma.

Photoallylation and photoreduction of cyclohexylidenepropanedinitrile by use of allyltrimethylsilane via photoinduced electron transfer: control of the product ratio depending on pK(a) values of additives.

Irradiation of an acetonitrile solution containing cyclohexylidenepropanedinitrile (1) and allyltrimethylsilane (2) in the presence of phenanthrene afforded two kinds of allylated products (3, 4) and a reduction product (5). The product ratio of 3, 4, and 5 dramatically changed depending on the pK(a) values of additives.

The Mouse Gene Expression Database (GXD).

The Gene Expression Database (GXD) is a community resource of gene expression information for the laboratory mouse. By combining the different types of expression data, GXD aims to provide increasingly complete information about the expression profiles of genes in different mouse strains and mutants, thus enabling valuable insights into the molecular networks that underlie normal development and disease. GXD is integrated with the Mouse Genome Database (MGD). Extensive interconnections with sequence databases and with databases from other species, and the development and use of shared controlled vocabularies extend GXD's utility for the analysis of gene expression information. GXD is accessible through the Mouse Genome Informatics web site at http://www.informatics.jax.org/ or directly at http://www.informatics.jax.org/menus/expression_menu. shtml.

Methylated Nepsilon-dansyl-L-lysine as a fluorogenic reagent for the chiral separation of carboxylic acids.

Dansyl amino acids having a free amino group and an asymmetric carbon atom were examined as a labeling reagent for chiral compounds containing carboxylic moieties to realize enantiomeric separation as well as fluorimetric determination. We tested dansyllysine by reacting it with (+)- and (-)-ibuprofen as a model carboxylic enantiomer. As the intramolecular carboxylic moiety of the dansyl amino acid interfered in the condensation reaction to form an amide bond with the carboxylic acid, the moiety was masked by methylation with trimethylsilyldiazomethane before the reaction. These derivatives were reacted with (+)- and (-)-ibuprofen and better enantiomeric resolution was achieved with methylated dansyllysine on a reversed-phase column. The derivatisation reaction was facilitated by the use of catalysts that are commonly employed in peptides synthesis. The reaction was completed within 5 min at room temperature when diethyl phosphorocyanidate was used. Due to the dansyl moiety, methylated dansyl-lysine enables a sensitive determination of ibuprofen with a fluorescence detector, in addition to the capability of enantiomer resolution. In tests, the detection limits for (+) and (-)-ibuprofen were 4 pmol per injection (S/N=3) at an excitation wavelength of 340 nm and an emission wavelength of 523 nm. Linear responses for the determination of (+) and (-)-ibuprofen in human urine were also demonstrated (r > or = 0.998) in the range from 10 to 1000 ng/ml. The precision and accuracy for urine samples spiked with (+)- and (-)-ibuprofen at 10, 100 and 1000 ng/ml were <10.1 and <14.6% (n=4), respectively.

Retinoic acid synthesis in mouse embryos during gastrulation and craniofacial development linked to class IV alcohol dehydrogenase gene expression.

Endogenous retinoic acid (RA) has been observed in vertebrate embryos as early as gastrulation, but the mechanism controlling spatiotemporal synthesis of this important regulatory molecule remains unknown. Some members of the alcohol dehydrogenase (ADH) family catalyze retinol oxidation, the rate-limiting step in RA synthesis. Here we have examined mouse embryos for the presence of endogenous RA and expression of ADH genes. RA was not detected in egg cylinder stage embryos but was detected in late primitive streak stage embryos. Detection of class IV ADH mRNA, but not class I or class III, coincided with the onset of RA synthesis, being absent in egg cylinder embryos but present in the posterior mesoderm of late primitive streak embryos. During neurulation, RA and class IV ADH mRNA were colocalized in the craniofacial region, trunk, and forelimb bud. Class IV ADH mRNA was detected in cranial neural crest cells and craniofacial mesenchyme as well as trunk and forelimb bud mesenchyme. The spatiotemporal expression pattern and enzymatic properties of class IV ADH are thus consistent with a crucial function in RA synthesis during embryogenesis. In addition, the finding of endogenous RA and class IV ADH mRNA in the craniofacial region has implications for the mechanism of fetal alcohol syndrome.

Digit induction by Hensen's node and notochord involves the expression of shh but not RAR-beta 2.

It is well established that Hensen's nodes can induce the formation of supernumerary digits after grafting into the anterior margin of the developing limb bud. The recent finding that distinct mesodermal cell populations are segregated within the node has made it possible to isolate different prospective cell types in an attempt to correlate digit-inducing ability with cell fate. We find that the prospective notochord cells contained within Hensen's node are able to induce supernumerary digits, whereas presumptive somite cells cannot. This early difference in inducing ability persists into later stages of development: epithelial somites are unable to induce while notochord from all lengths of the neuraxis continues to induce. Using probes to retinoic acid receptor-beta 2 and sonic hedgehog (shh) we find no evidence to support the idea that inducing tissues generate extra digits by releasing retinoic acid into adjacent limb tissue but find that the inducing ability of a tissue correlates with its expression of shh.

Reciprocal changes in Hox D13 and RAR-beta 2 expression in response to retinoic acid in chick limb buds.

The finding that retinoic acid (RA) has major effects on pattern formation in developing chick limbs has led to the conclusion that RA plays a central role in the normal development of the limb. In addition, it has been demonstrated that treatment with exogenous RA leads to changes in expression of homeobox-containing Hox complex genes within the developing limb. This has been used to argue that, since RA activates 5' Hox D complex genes in the same temporal and spatial sequence as in normal development, Hox genes might be regulated by RA during normal limb development. In this study, we further examine the temporal and spatial changes in expression of Hox D13 and RAR-beta 2 transcripts in wing buds in response to local application of RA in vivo. We confirm reports that RAR-beta 2 expression is induced early and locally by RA. However, we find that the effect of RA on Hox D13 gene expression at the distal end of wing buds at stage 25/26 is downregulation of transcript expression. Furthermore, we find that activation of ectopic Hox D13 expression in response to implantation of RA beads along the anterior margin at stage 20/21 is indirect. Finally, the effects of RA exposure on Hox D13 expression appear to directly correlate with effects on the pattern of distal skeletal elements.

Regeneration of HoxD expression domains during pattern regulation in chick wing buds.

The expression domains of genes located at the 5' end of the HoxD (formerly Hox-4) complex appear to correlate with pattern along both the proximal-distal (PrDi) and the anterior-posterior (AP) axes of the developing limb bud, and it has been suggested that the HoxD gene products are involved in the specification of positional information during limb development. The apical ectodermal ridge is required for limb outgrowth and is thought to influence mesodermal cells at the distal end of the limb bud in a region within which patterning events occur. In this paper, we examine the expression of 5' HoxD genes during PrDi pattern regulation in chick wing buds. In limbs undergoing pattern regulation, we demonstrate that the domains of HoxD11 and HoxD13 gene expression are "regenerated" within 24 hr of removal of the distal mesenchyme. In contrast, in limbs which will not form distal structures, HoxD13 expression becomes reduced.

Retinoic acid respecifies limb bud cells in vitro.

Retinoic acid (RA) is known to have dramatic effects on limb pattern formation and has been shown to exert its effects on limbs by converting anterior limb bud cells into cells with posterior positional properties. In this study we find that dissociated posterior limb bud cells from chick and mouse embryos cultured at high density (micromass cultures) are able to stimulate the formation of supernumerary digits when grafted into developing wing buds and that the positional identity of both chick and mouse limb bud cells can be maintained for finite periods of time in vitro. Furthermore, using this assay system we have tested whether anterior cells from mouse and chick limb buds can be converted into cells with posterior identity by exposure to RA in vitro. We find that anterior limb bud cells acquire posterior properties after culture in the presence of RA.

Effects of localized application of transforming growth factor beta 1 on developing chick limbs.

The effects of exogenous transforming growth factor beta (TGF-beta) on chick limb development in vivo were studied by implanting carriers of TGF-beta 1 into developing wing buds. Agarose beads were soaked in solutions containing TGF-beta 1 and implanted into wing buds at stages 18 to 27. Localized application of TGF-beta 1 to distal regions of the wing bud caused specific skeletal elements in the limb to be reduced or absent. The particular proximal-distal limb element affected depended on the stage at which the bead was implanted. Position of the bead in the anterior-posterior axis also influenced the pattern of affected structures. Experiments in which TGF-beta 1 beads were implanted and then removed at 24- and 48-hr intervals indicate that there are specific periods during which a skeletal element appears to be sensitive to the effects of exogenous TGF-beta 1. In a few cases, beads placed in proximal positions in later staged limbs resulted in formation of ectopic cartilage near the bead. These results suggest that exposure to exogenous TGF-beta 1 in vivo influences the development of skeletal elements in the chick limb in a stage- and position-dependent manner.

A comparison of hemostatic agents in microvascular surgery.

The hemostatic abilities of oxidized cellulose, gelatin powder, microfibrillar collagen, and expanded polytetrafluoroethylene cuffs were compared on anastomoses of the femoral vessels in rats. The intraoperative hemostatic time and patency were recorded. Vessels that were patent intraoperatively were examined at various intervals postoperatively to determine patency and to detect the development of abnormalities, and specimens for examination under a scanning electron microscope were obtained. Microfibrillar collagen was found to be superior in maintaining patency and in ease of use. The PTFE cuffs demonstrated the shortest hemostatic times, but they produced distortion of the vessels several weeks after surgery. The authors conclude that microfibrillar collagen is the best overall hemostatic agent and have continued to use it experimentally and clinically with no complications.