Interaction of Wnt and caudal-related genes in zebrafish posterior body formation

AbstractAlthough Wnt signaling plays an important role in body patterning during early vertebrate embryogenesis, the mechanisms by which Wnts control the individual processes of body patterning are largely unknown. In zebrafish, wnt3a and wnt8 are expressed in overlapping domains in the blastoderm margin and later in the tailbud. The combined inhibition of Wnt3a and Wnt8 by antisense morpholino oligonucleotides led to anteriorization of the neuroectoderm, expansion of the dorsal organizer, and loss of the posterior body structure–a more severe phenotype than with inhibition of each Wnt alone–indicating a redundant role for Wnt3a and Wnt8. The ventrally expressed homeobox genes vox, vent, and ved mediated Wnt3a/Wnt8 signaling to restrict the organizer domain. Of posterior body-formation genes, expression of the caudal-related cdx1a and cdx4/kugelig, but not bmps or cyclops, was strongly reduced in the wnt3a/wnt8 morphant embryos. Like the wnt3a/wnt8 morphant embryos, cdx1a/cdx4 morphant embryos displayed complete loss of the tail structure, suggesting that Cdx1a and Cdx4 mediate Wnt-dependent posterior body formation. We also found that cdx1a and cdx4 expression is dependent on Fgf signaling. hoxa9a and hoxb7a expression was down-regulated in the wnt3a/wnt8 and cdx1a/cdx4 morphant embryos, and in embryos with defects in Fgf signaling. Fgf signaling was required for Cdx-mediated hoxa9a expression. Both the wnt3a/wnt8 and cdx1a/cdx4 morphant embryos failed to promote somitogenesis during mid-segmentation. These data indicate that the cdx genes mediate Wnt signaling and play essential roles in the morphogenesis of the posterior body in zebrafish

Fine ice particles formed at dew point of -70℃

第2回極域科学シンポジウム/第34回気水圏シンポジウム 11月14日（月） 統計数理研究所 セミナー室

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

Electron Beam Lithography for Large Area Patterning 1: Development of Large Field Deflection E-Beam Lithography System

A novel electron beam system has been designed and developed specifically for large area patterning of electronic devices such as printed wiring boards. The prototyped system features a large field deflection, high scanning speed and stably focused beam in the large field. An electron gun with a LaB5 flat cathode was used by operating at 1750-1800 K. The electron beam column provides an electron probe of less than 40 μm in diameter with a current of 50 μA at 60 kV. Fast and large field deflections by a magnetic deflection system enables an area of 104 mm x 104 mm to be covered. The scanning speed can range up to 254 m/s. Particular attention was paid to the materials and shapes of the optics column to minimize the influence of eddy currents from the point of view of controlling the dynamic behavior of beam deflection. It is confirmed that the system can provide accurate beam deflection within a ±20 μm (3) tolerance for the quite large field of 52 mm x 52 mm

Dynamic effects of the Nuss procedure on the spine in asymmetric pectus excavatum

ObjectiveThis study aimed to elucidate dynamic effects of the Nuss procedure on the spine in the treatment of patients with pectus excavatum with asymmetric thoraces.MethodsTwenty-five patients with pectus excavatum who underwent the Nuss procedure were categorized into 4 groups by preoperative morphology of the spine and thoracic asymmetry. In group 1 (n = 8), the right side of the thorax was concave and the spine bowed to the right. In group 2 (n = 4), the right side of the thorax was concave and the spine bowed to the left. In group 3 (n = 5), the left side of the thorax was concave and the spine bowed to the right. In group 4 (n = 8), the left side of the thorax was concave and the spine bowed to the left. With computed tomographic data, finite-element models were produced to simulate each patient's thorax. Thereafter, dynamic response patterns of the spine to the Nuss procedure were examined. Validity of these biomechanical findings was verified by referring to clinical outcomes.ResultsIn group 1 and group 4 models, deformed spines were straightened; in group 2 and group 3 models, spinal bowing increased. These biomechanical findings were compatible with clinical evaluations.ConclusionsPerformance of the Nuss procedure for asymmetric pectus excavatum exerts dynamic influence on the spine. Response patterns of the spine are predictable from morphologic relationships between the asymmetric patterns of the anterior thoracic wall and the spine

Effect of intracoronary thrombectomy on 30-day mortality in non-diabetic patients with acute hyperglycemia after acute myocardial infarction

SummaryBackgroundThere is limited evidence about useful therapeutic interventions for patients with acute hyperglycemia (AH) after acute myocardial infarction (AMI).MethodsWe studied 2433 consecutive non-diabetic AMI patients who underwent percutaneous coronary intervention (PCI) within 24h after the onset. Patients were divided into two groups according to the presence or absence of AH (admission serum glucose level ≥11.1mmol/l). We assessed the association between intracoronary thrombectomy and the clinical outcome in AMI patients with AH.ResultsPatients with AH had more risk factors than those without AH. The 30-day mortality rate of patients with AH was significantly higher than that of those without (11.7% vs 1.7%, p<0.001). Among patients with AH, the 30-day mortality rate was significantly lower for those with intracoronary thrombectomy than those without it (4.9% vs 17.2%, p=0.004). Among patients without AH, however, the 30-day mortality rate was similar between those with and without intracoronary thrombectomy (1.5% vs 1.9%, p=NS). Multivariate analysis showed that intracoronary thrombectomy was associated with an improved 30-day mortality rate for patients with AH (hazard ratio: HR 0.184, 95% CI 0.057–0.598, p=0.005).ConclusionsIn AMI patients with AH, intracoronary thrombectomy prior to PCI might improve the 30-day mortality rate

Singlet oxygen -derived nerve growth factor exacerbates airway hyperresponsiveness in a mouse model of asthma with mixed inflammation

Background: Refractory asthma, which is caused by several factors including neutrophil infiltration is a serious complication of bronchial asthma. We previously reported that nerve growth factor (NGF) is involved in AHR. NGF-derived induction of hyperalgesia is dependent on neutrophils; however, this relationship remains unclear in respiratory disease. In this study, we examined the roles of neutrophils and NGF in refractory asthma. Methods: Using intranasal house dust mite sensitization, we established a mouse model of asthma with mixed inflammation (Mix-in). AHR, NGF production and hyperinnervation of the lungs were examined with or without different inhibitory treatments. The levels of the singlet oxygen markers, 10- and 12-(Z,E)-hydroxyoctadecadienoic acids (HODE) in the lungs, were measured by liquid chromatography-tandem mass spectrometry. An in vitro experiment was also performed to evaluate the direct effect of singlet oxygen on NGF production. Results: NGF production and hyperinnervation were higher in Mix-in mice than in conventional eosinophilic-asthmatic mice and were positively correlated with AHR. Asthmatic parameters were inhibited by NGF neutralizing Abs and myeloperoxidase (MPO) inhibition. The 10- and 12-(Z,E)-HODEs levels were increased in the lungs and were positively correlated with MPO activity and NGF production. NGF was produced by bronchial epithelial cells in vitro upon stimulation with singlet oxygen. Conclusions: Our findings suggest that neutrophil MPO-derived singlet oxygen induces increased NGF production, leading to AHR and 10- and 12-(Z,E)-HODEs production. These findings may help to develop new therapies targeting this mechanism and to establish a new biomarker for non-type 2 and refractory asthma

Magnetostriction studies up to megagauss fields using fiber Bragg grating technique

We here report magnetostriction measurements under pulsed megagauss fields using a high-speed 100 MHz strain monitoring system devised using fiber Bragg grating (FBG) technique with optical filter method. The optical filter method is a detection scheme of the strain of FBG, where the changing Bragg wavelength of the FBG reflection is converted to the intensity of reflected light to enable the 100 MHz measurement. In order to show the usefulness and reliability of the method, we report the measurements for solid oxygen, spin-controlled crystal, and volborthite, a deformed Kagom\'{e} quantum spin lattice, using static magnetic fields up to 7 T and non-destructive millisecond pulse magnets up to 50 T. Then, we show the application of the method for the magnetostriction measurements of CaV$_{4}$O$_{9}$, a two-dimensional antiferromagnet with spin-halves, and LaCoO$_{3}$, an anomalous spin-crossover oxide, in the megagauss fields.Comment: 9pages, 6 figures, Conference proceedings for MegaGauss16 at Kashiwa, Japan in Sept. 201