Terahertz absorption spectra of L-, D-, and DL-alanine and their application to determination of enantiometric composition

Absorption spectra of polycrystalline L-, D-, and DL-alanine have been measured by the terahertz time domain spectroscopy (THz-TDS)in the frequency range from 10 to 90 cm−1 at room temperature. We observed several absorption bands, which have the large difference between enantiomers (L- and D-alanine)and the racemic compound (DL-alanine)in their peak frequencies. This obvious difference shows that the THz absorption bands are strikingly sensitive to the crystal structures. This result indicates that the THz-TDS can be used for distinguishing between the enantiomers and the racemic compound. We propose and demonstrate a method to determine the enantiometric composition of amino acids from the THz absorption spectra

N,N’-Bis(2-chloroethyl)-N-nitrosourea (BCNU)-induced Apoptosis of Neural Progenitor Cells in the Developing Fetal Rat Brain

N,N’-bis(2-chloroethyl)-N-nitrosourea (BCNU) is one of the major drugs used in chemotherapy against malignant gliomas due to its effects, such as induction of bifunctional alkylation of DNA and formation of interstrand DNA cross-linkages, and induces cortical malformations in the fetal and neonatal rat brain. In this study, pregnant rats were treated with 7.5 mg/kg of BCNU on gestational day 13 (GD 13), and their fetuses were collected from 12 to 72 hours after BCNU treatment in order to examine the timecourses of morphological and immunohistochemical changes in neural progenitor cells in the developing brain. The number of pyknotic cells in the telencephalon peaked at 24 h and then gradually decreased until 72 h. The majority of these pyknotic cells were positive for cleaved caspase-3, a key executioner of apoptosis. The pyknotic cells showed the ultrastructural characteristics of apoptosis. The number of p53-positive cells began to increase prior to the appearance of apoptotic cells and p21-positive cells. The number of phosphorylated-histone H3-positive cells (mitotic cells) decreased from 24 to 36 h. The number of Iba1-positive cells (microglial cells) in the telencephalon increased from 12 to 48 h. These results suggest that BCNU induces p53-dependent apoptosis and reduces proliferative activity, resulting in reduction of the weight of the telencephalon and the thickness of the telencephalic wall in the fetal brain. This study will help to clarify the mechanisms of BCNU-induced fetal brain toxicity

Tracing of Afferent Connections in the Zebrafish Cerebellum Using Recombinant Rabies Virus

The cerebellum is involved in some forms of motor coordination and learning, and in cognitive and emotional functions. To elucidate the functions of the cerebellum, it is important to unravel the detailed connections of the cerebellar neurons. Although the cerebellar neural circuit structure is generally conserved among vertebrates, it is not clear whether the cerebellum receives and processes the same or similar information in different vertebrate species. Here, we performed monosynaptic retrograde tracing with recombinant rabies viruses (RV) to identify the afferent connections of the zebrafish cerebellar neurons. We used a G-deleted RV that expressed GFP. The virus was also pseudotyped with EnvA, an envelope protein of avian sarcoma and leucosis virus (ALSV-A). For the specific infection of cerebellar neurons, we expressed the RV glycoprotein (G) gene and the envelope protein TVA, which is the receptor for EnvA, in Purkinje cells (PCs) or granule cells (GCs), using the promoter for aldolase Ca (aldoca) or cerebellin 12 (cbln12), respectively. When the virus infected PCs in the aldoca line, GFP was detected in the PCs’ presynaptic neurons, including GCs and neurons in the inferior olivary nuclei (IOs), which send climbing fibers (CFs). These observations validated the RV tracing method in zebrafish. When the virus infected GCs in the cbln12 line, GFP was again detected in their presynaptic neurons, including neurons in the pretectal nuclei, the nucleus lateralis valvulae (NLV), the central gray (CG), the medial octavolateralis nucleus (MON), and the descending octaval nucleus (DON). GFP was not observed in these neurons when the virus infected PCs in the aldoca line. These precerebellar neurons generally agree with those reported for other teleost species and are at least partly conserved with those in mammals. Our results demonstrate that the RV system can be used for connectome analyses in zebrafish, and provide fundamental information about the cerebellar neural circuits, which will be valuable for elucidating the functions of cerebellar neural circuits in zebrafish

CYK4 relaxes the bias in the off-axis motion by MKLP1 kinesin-6

Centralspindlin, a complex of the MKLP1 kinesin-6 and CYK4 GAP subunits, plays key roles in metazoan cytokinesis. CYK4-binding to the long neck region of MKLP1 restricts the configuration of the two MKLP1 motor domains in the centralspindlin. However, it is unclear how the CYK4-binding modulates the interaction of MKLP1 with a microtubule. Here, we performed three-dimensional nanometry of a microbead coated with multiple MKLP1 molecules on a freely suspended microtubule. We found that beads driven by dimeric MKLP1 exhibited persistently left-handed helical trajectories around the microtubule axis, indicating torque generation. By contrast, centralspindlin, like monomeric MKLP1, showed similarly left-handed but less persistent helical movement with occasional rightward movements. Analysis of the fluctuating helical movement indicated that the MKLP1 stochastically makes off-axis motions biased towards the protofilament on the left. CYK4-binding to the neck domains in MKLP1 enables more flexible off-axis motion of centralspindlin, which would help to avoid obstacles along crowded spindle microtubules

研究資料室中央資料庫収蔵資料の公開に向けての取り組みと課題

国立国語研究所 研究情報発信センター 非常勤研究員国立国語研究所 研究情報発信センター 非常勤研究員Adjunct Researcher, Center for Research Resources, NINJALAdjunct Researcher, Center for Research Resources, NINJAL国立国語研究所はこれまで方言，語彙，日本語教育等の様々な調査や研究プロジェクトを実施し，その過程で数多くの研究資料を収集・作成してきた。これらの貴重な研究資料は現在研究資料室に収蔵されているが，国立国語研究所の使命を考えれば研究所内外に広く公開し，日本語の研究・教育活動のために役立てられることが望ましい。本稿では，研究資料室中央資料庫に収蔵されている主に紙媒体の研究資料について，公開に向けて行った準備と，その過程で生じた課題について述べる。The National Institute for Japanese Language and Linguistics (NINJAL) has promoted many collaborative research studies, in areas such as dialects, vocabularies, and Japanese language education, and in the process, has collected and preserved research materials at the Center for Research Resources. Considering NINJAL\u27s mission, it is desirable to open research materials to the public for deserving Japanese language research and education. This paper describes an approach and issue in relation to the public opening of paper research materials

752-5 Dose the “No Reflow” Phenomenon Immediately After Successful Direct Angioplasty Always Indicate Poor Functional Recovery?

Experimental studies have demonstrated that “no reflow” phenomenon after prolonged coronary occlusion indicates advanced myocardial necrosis. In clinical settings, previous studies have suggested that presence of “no reflow” phenomenon following thrombolysis is a useful predictor of poor functional recovery. To test the hypothesis that “no reflow” phenomenon is closely related to the recovery of regional wall motion in a reperfused AMI, we studied 42 myocardial segments in 18 patients with AMI who achieved successful direct angioplasty. MCE was performed immediately after direct angioplasty and repeated 2 weeks later with intracoronary injection of sonicated albumin. Contrast effect in each segment was scored as 0–2 (0, 1 and 2 denoting no, partial and homogeneous contrast effect, respectively). 2D-Echo was performed at day-1 and repeated 1 month later. Wall motion (WM) by 2D-Echo was scored as 0–3 (scores of 0 to 3 indicating dys/akinetic segments to normal. respectively). The relation between mean WM and MCE score immediately after direct angioplasty was as follows:MCE scoreday-l WM1 Month WMChange2 (n=20)1.1±0.8**1.7±0.7*,#0.6±0.91 (n=7)0.6±0.8*1.4±1.4#0.9±0.90 (n=15)0.0±0.00.8±1.0##0.8±1.0*p<0.01**p<0.001 vs. MCE score=0#p<0.05##p<0.01 vs. day-l WMSeven of 15 segments (47%) with MCE score=0 immediately after direct angioplasty showed functional recovery at 1 month later, and 6 of the 7 segments (86%) showed contrast enhancement at 2 weeks later restudy.ConclusionAbout one half of segments which showed no contrast enhancement immediately after direct angioplasty showed functional recovery at 1 month later, and the majority of these segments showed contrast enhancement at 2 weeks later restudy. The “no reflow” phenomenon by MCE immediately after successful direct angioplasty dose not always indicate poor functional recovery