ジゴキシゲニン標識プローブを用いたBeet Necrotic Yellow Vein Virus RNA の検出

Complementary DNA (cDNA) clones corresponding to each of five distinct RNA species of beet necrotic yellow vein (BNYVV) were synthesized and identified. The sizes of each cDNA clone for RNAs 1,2,3,4 and 5 molecules were 3.0, 1.7, 1.8, 1.5 and 1.4 kbp, respectively. cDNA inserts to RNA 2 were covered at a part of the 3'regions, and those of RNAs 3,4 and 5 were almost full-length. The plasmids containing each of cDNA inserts were labeled with digoxigenin by the random priming method. Northern blot hybridization tests showed that individual probes hybridized specially to each of the five RNAs. Good results were obtained with 1 to 10 ng of RNA as a mixture of five RANs, but the probe to RNA 3, RNA 4 or RNA 5 gave a weak signal with hererologous RNAs when more than 10 ng RNA was used. In dot blot hybridization, the limit of detection was about 10 pg RNA, but if a higher content of RNA was spotted, cross reaction occurred using heterologous RNAs. For laboratory and field isolates of BNYVV, each of RNAs 3,4 and 5 was easily detected by Northern blot hybridization in total nucleic acids extracted from Tetragonia expansa leaves inoculated mechanically, but not from roots of sugar-beet plants inoculated by the fungus Polymyxa betae. However, satisfactory results were obtained with partially purified or concentrated preparations from roots. These findings indicate that the digoxigenin-labeled probes are useful for the identification and detection of RNAs contained in field and laboratory isolates of BNYVV.テンサイそう根ウイルス（Beet nectotic yellow vien virus,BNYVV）に含まれる5種類のRNAのcDNAをクローニングした。得られたcDNAクローンのサイズは次の通りであった：RNA－1、3.0kbp；RNA－2、1.7kbp；RNA-3、1.8kbp；RNA－4、1.5kbp；RNA－5、1.4kbp。RNA-1とRNA-2由来のcDNAクローンは、それぞれ全ゲノムの3’末端側45％と35％を占めていた。RNA-3、RNA－4およびRNA－5からのcDNAクローンはほぼ全長であった。各cDNAクローンをジゴキシゲニン標識し、Northern blot hybridization でウイルスRNAの検出条件を検討した結果、試料はホルマリン・ホルムアルデミド変性、プローブのソニケーション、プローブ量は50ng/ml,ホルムアミド存在下の温度は42℃で行うのが最適であった。BNYVVの4～5種のRNAを検出するためには、全RNA量として1～10ngが適当であった。全RNA量が20ng以上では、RNA-3、RNA-4およびRNA-5の間でお互いにわずかに反応が認められた。この原因は、プローブの3’末端にみられる共通配列によることが確かめられた。Dot blot hybridization によるウイルスRNAの検出感度は、10pgであったが、試料の濃度が高くなるにつれて、RNA-3、RNA-4およびRNA-5の間でお互いに反応が認められた。各種BNYVV分離株を用いて、それらに存在するRNA組成とサイズをNorthern blot hybridization により解析したところ、検定植物ツルナ接種葉の場合には、直接全核酸を調整した試料からウイルスRNAの検出が可能であったが、Polymyxa betae 菌で接種されたテンサイの根の場合には、ウイルス濃度に応じて部分純化または濃縮した試料から核酸を調整する必要があった。このような条件下で、ほ場および室内分離株に存在するBNYVV RNA種およびそれらの欠失変異株を確実に検出することができた

Changes in Knowledge Levels through Lectures on Radiotherapy to Nursing Students in Japan

Background:Recently, the number of patients choosing radiotherapy is increasing in Japan. It is necessary for even nurses to have knowledge of radiotherapy. We measured the degree of awareness with a lecture on how a nursing student can obtain knowledge of radiotherapy.Materials and methods: Forty nursing students who were in their third year and preparing for their national nursing certification in two months were recruited. The 90-min lecture covered a range of topics from general theories to specific concepts. The students were not informed that they would be required to fill a survey at the end of the lecture. This was to avoid bias that could occur from knowing in advance that a survey would be conducted.Results: The following items below were particularly remarkable. The distinction between the radiologist and the radiation oncologist. The radiotherapy is a local therapy. The pain from bone metastasis could be relieved.Conclusion: Students had very little knowledge on radiotherapy before the lecture. the results of this study indicated the need to increase the number of lectures on radiotherapy for nursing as well medical students. As the number of radiotherapy patients increases

Pathogenetic roles of beet necrotic yellow vein virus RNA5 in the exacerbation of symptoms and yield reduction, development of scab‐like symptoms, and Rz1‐resistance breaking in sugar beet

Beet necrotic yellow vein virus (BNYVV) generally has a four‐segmented positive‐sense RNA genome (RNAs 1–4), but some European and most Asian strains have an additional segment, RNA5. This study examined the effect of RNA5 and RNA3 on different sugar beet cultivars using a Polymyxa‐mediated inoculation system under field and laboratory conditions. In field tests, the degree of sugar yield served as an index for assessing the virulence of BNYVV strains. Japanese A‐II type isolates without RNA5 caused mostly 15%–90% sugar yield reductions, depending on the susceptibility of sugar beet cultivars, whereas the isolates with RNA5 induced more than 90% yield losses in the seven susceptible cultivars, but small yield losses in one Rz1‐resistant and Rizor cultivars. However, a laboratory‐produced isolate containing RNA5 but lacking RNA3 caused higher yield losses in Rizor than in susceptible plants, and induced scab‐like symptoms on the root surface of both susceptible and resistant plants. In laboratory tests, A‐II type isolates without RNA5 had low viral RNA accumulation levels in roots of Rizor and Rz1‐resistant plants at early stages of infection, but in the presence of RNA5, viral RNA3 accumulation levels increased remarkably. This increased RNA3 accumulation was not observed in roots of the WB42 accession with the Rz2 gene. In contrast, the presence of RNA3 did not affect RNA5 accumulation levels. Collectively, this study demonstrated that RNA5 is involved in the development of scab‐like symptoms and the enhancement of RNA3 accumulation, and suggests these characteristics of RNA5 are associated with Rz1‐resistance breaking

Development of an Oropharyngeal Scope with an Integrated Tongue Depressor: NTOP2013 Study

The oropharynx is examined with a light source such as an electric light, a penlight, or a forehead mirror based on an acquired visual field using a tongue depressor. However, it is extremely difficult to obtain objective and reproducible images of tissue within the pharynx required in recent years with these methods, and insufficient progress in the examination tools has been made. There is an increasing need to develop a method for display during oropharyngeal examination. We conducted the present study to develop a novel oropharyngeal endoscope as an objective observation method

Validation of MALDI-TOF MS devices in reanalysis of unidentified pathogenic bacteria detected in blood cultures

In hospital microbial laboratories, morphological and biochemical analyses are performed to identify pathogenic microbes;however, these procedures lack rapidity and accuracy. Recently, Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF MS) has been clinically utilized, and is expected to enable rapid and accurate microbial identification. We aimed to validate two MALDI-TOF MS devices available in Japan: the VITEK-MS (BioMérieux) and the Microflex LT (Bruker Daltonics). Clinically isolated bacteria, 100 samples in all, detected in blood cultures but incompletely identified by conventional procedures, were reanalyzed using the two devices. The VITEK-MS and Microflex LT, respectively, identified 49% (49/100) and 80% (80/100) of the tested bacteria at the species level, as well as 96% (96/100) and 95% (95/100) at the genus level. Among those reidentified strains, 26% (26/100) at the species level and 88% (88/100) at the genus level were concordant with each other, though three strains were unmatched. Moreover, four bacterial strains were unable to be identified using the VITEK-MS, versus five using the Microflex LT. MALDI-TOF MS devices can provide more rapid and accurate bacterial identification than ever before;however, the characteristics of each system were slightly different;therefore, it is necessary to understand the difference in performance of MALDI-TOF MS models

緩和ケアにおける人材育成研修の成果と課題 : 修了生の看護実践・自己の姿勢への活用

A県における緩和ケアの研修を修了した修了生173名に、研修後の看護実践と自己の行動の変化について質問紙調査を行った。その結果、身体症状の緩和や傾聴、日常生活への支援に関する看護実践、自己の姿勢・態度の変容に活かしやすく、精神症状の緩和、遺族ケア、意思決定、地域連携に関する看護実践には活かしにくかった。一方、スキルアップの動機づけにもなっていた。今後は、難しいと感じる分野の研修内容・方法の検討、モチベーションの維持・向上のための環境調整やフォローアップの機会が必要であることが明らかになった

修士論文要旨 : F. 臨床心理学研究領域

departmental bulletin pape

Mechanistic Study on Gold-Like Luster Development of Solution-Cast Oligo(3-methoxythiophene) Film

Solution-cast coating films of perchlorate-doped oligo(3-methoxythiophene) exhibited a gold-like luster similar to that of metallic gold despite the involvement of no metals. However, the development mechanism of the luster remains ambiguous. To understand the mechanism, we performed scanning electron microscopic analysis, variable-angle spectral reflectance measurements, and ellipsometry measurements on ClO4−-doped oligo(3-methoxythiophene) cast film with a gold-like luster. The results revealed that the lustrous color of the film was not induced by the submicron-sized regular structures (structural color), nor by the high-density free electrons (reflective response based on Drude model), but by the large optical constants (refractive index and extinction coefficient) of the film, as speculated previously