Sets of RNA Repeated Tags and Hybridization-Sensitive Fluorescent Probes for Distinct Images of RNA in a Living Cell

BACKGROUND: Imaging the behavior of RNA in a living cell is a powerful means for understanding RNA functions and acquiring spatiotemporal information in a single cell. For more distinct RNA imaging in a living cell, a more effective chemical method to fluorescently label RNA is now required. In addition, development of the technology labeling with different colors for different RNA would make it easier to analyze plural RNA strands expressing in a cell. METHODOLOGY/PRINCIPAL FINDINGS: Tag technology for RNA imaging in a living cell has been developed based on the unique chemical functions of exciton-controlled hybridization-sensitive oligonucleotide (ECHO) probes. Repetitions of selected 18-nucleotide RNA tags were incorporated into the mRNA 3'-UTR. Pairs with complementary ECHO probes exhibited hybridization-sensitive fluorescence emission for the mRNA expressed in a living cell. The mRNA in a nucleus was detected clearly as fluorescent puncta, and the images of the expression of two mRNAs were obtained independently and simultaneously with two orthogonal tag-probe pairs. CONCLUSIONS/SIGNIFICANCE: A compact and repeated label has been developed for RNA imaging in a living cell, based on the photochemistry of ECHO probes. The pairs of an 18-nt RNA tag and the complementary ECHO probes are highly thermostable, sequence-specifically emissive, and orthogonal to each other. The nucleotide length necessary for one tag sequence is much shorter compared with conventional tag technologies, resulting in easy preparation of the tag sequences with a larger number of repeats for more distinct RNA imaging

Methods for Estimating Effective Diffusivity of Substrate and Kinetic Parameters of Immobilized Enzyme

Two methods were presented for estimating simultaneously the kinetic parameters in the Michaelis-Menten equation, Kₘ and Vₘₐₓ, and the intraparticle effective diffusivity of substrate, Dₑᴀ, from the results of the transient changes in a batch reactor. The methods were applied to the estimation of the Kₘ and Vₘₐₓ values of α-chymotrypsin immobilized into firebrick particles or acrylamide gel, and the Dₑᴀ values of substrate through the supports. The experimental data of conversions both in the batch and tubular reactors were found to be calculated successfully by using the kinetic and transport parameters estimated by the proposed methods

Development of an autonomous FM-CW ionospheric observation system based on reinforcement learning

第150回地球電磁気・地球惑星圏学会（SGEPSS）総会および講演会, 2021年10月31日-11月4

Seasonal and long-term variations in the Schumann Resonance observed at Kuju Japan

第150回地球電磁気・地球惑星圏学会（SGEPSS）総会および講演会, 2021年10月31日-11月4

Cryptococcus gattii Genotype VGIIa Infection in Man, Japan, 2007

We report a patient in Japan infected with Cryptococcus gattii genotype VGIIa who had no recent history of travel to disease-endemic areas. This strain was identical to the Vancouver Island outbreak strain R265. Our results suggest that this virulent strain has spread to regions outside North America

Large-scale mapping observations of the CI(3P1-3P0) and CO(J=3-2) lines toward the Orion A molecular cloud

Large scale mapping observations of the 3P1-3P0 fine structure transition of atomic carbon (CI, 492 GHz) and the J=3-2 transition of CO (346 GHz) toward the Orion A molecular cloud have been carried out with the Mt. Fuji submillimeter-wave telescope. The observations cover 9 square degrees, and include the Orion nebula M42 and the L1641 dark cloud complex. The CI emission extends over almost the entire region of the Orion A cloud and is surprisingly similar to that of 13CO(J=1-0).The CO(J=3-2) emission shows a more featureless and extended distribution than CI.The CI/CO(J=3-2) integrated intensity ratio shows a spatial gradient running from the north (0.10) to the south (1.2) of the Orion A cloud, which we interpret as a consequence of the temperature gradient. On the other hand, the CI/13CO(J=1-0) intensity ratio shows no systematic gradient. We have found a good correlation between the CI and 13CO(J=1-0) intensities over the Orion A cloud. This result is discussed on the basis of photodissociation region models.Comment: Text file is 13 pages long, and 3 figure files (pdf format). NRO Report No. 508 (1999). University of Tokyo, Resceu 41/9

Management of Multiple Primary Lung Cancer in Patients with Centrally Located Early Cancer Lesions

BackgroundPatients with centrally located early lung cancer (CLELC) are often heavy smokers with a considerably high risk of multiple primary lung cancer (MPLC) lesions; treatment strategies for such patients must preserve the cardiopulmonary function.MethodsBetween July 2004 and July 2008, patients with CLELC underwent photodynamic therapy (PDT) using NPe6, second-generation photosensitizer at Tokyo Medical University Hospital. Among these patients, we retrospectively analyzed MPLC, which was treated by surgery plus PDT or PDT alone and examined the effectiveness of PDT, and we propose a treatment strategy for patients with MPLC.ResultsA total of 64 patients with CLECL received NPe6-PDT, and MPLCs were found in 22 patients (34.4%) using sputum cytology and a bronchoscopical examination using autofluorescence bronchoscopy. Among these 22 patients, 10 patients underwent surgery for primary lung cancer and underwent NPe6-PDT for the treatment of secondary primary CLELC, one patient underwent PDT for CLELC as a primary lesion followed by an operation for peripheral-type lung cancer as a secondary primary lesion, and 11 patients underwent PDT alone for MPLC lesions (28 lesions) that were roentgenographically occult lung cancers. Among these 22 patients with MPLC including peripheral-type lung cancers, which were resected by surgery, all 39 CLELC lesions exhibited a complete response after PDT, and all patients were alive.ConclusionsFor patients with lung cancer with a long-term history of smoking, careful follow-up examinations after surgical resection are needed considering the incidence of metachronous primary lung cancers. PDT can play an important role for the treatment strategy for MPLC

Comparative cytology of stromal tumors of the gastrointestinal tract

The cytologic features of four cases of stromal tumors arising in the gastrointestinal tract are presented here. Two benign schwannomas of the stomach showed characteristic findings of spindle cells within a background containing abundant mature lymphcytes. A benign gastrointestinal stromal tumor (GIST) of the duodenum demonstrated small clusters composed of spindle cells with uniform and cigar-shaped nuclei. A malignant GIST metastatic to the liver was composed of isolated plump spindle cells with hyperchromatic nuclei with multiple small nucleoli. The diagnosis and classification of a gastric or intestinal stromal tumor is possible based on the cytologic findings