A trial of somatic gene targeting in vivo with an adenovirus vector

BACKGROUND: Gene targeting in vivo provides a potentially powerful method for gene analysis and gene therapy. In order to sensitively detect and accurately measure designed sequence changes, we have used a transgenic mouse system, MutaMouse, which has been developed for detection of mutation in vivo. It carries bacteriophage lambda genome with lacZ(+ )gene, whose change to lacZ-negative allele is detected after in vitro packaging into bacteriophage particles. We have also demonstrated that gene transfer with a replication-defective adenovirus vector can achieve efficient and accurate gene targeting in vitro. METHODS: An 8 kb long DNA corresponding to the bacteriophage lambda transgene with one of two lacZ-negative single-base-pair-substitution mutant allele was inserted into a replication-defective adenovirus vector. This recombinant adenovirus was injected to the transgenic mice via tail-vein. Twenty-four hours later, genomic DNA was extracted from the liver tissue and the lambda::lacZ were recovered by in vitro packaging. The lacZ-negative phage was detected as a plaque former on agar with phenyl-beta-D-galactoside. RESULTS: The mutant frequency of the lacZ-negative recombinant adenovirus injected mice was at the same level with the control mouse (~1/10000). Our further restriction analysis did not detect any designed recombinant. CONCLUSION: The frequency of gene targeting in the mouse liver by these recombinant adenoviruses was shown to be less than 1/20000 in our assay. However, these results will aid the development of a sensitive, reliable and PCR-independent assay for gene targeting in vivo mediated by virus vectors and other means

Clinicopathologic Analysis of Sinonasal Inverted Papilloma, with Focus on Human Papillomavirus Infection Status

Sinonasal inverted papilloma (SNIP) can recur; however, the factors related to tumor recurrence remain unclear. This study aimed to analyze risk factors, including human papillomavirus (HPV) infection, as well as other factors associated with SNIP recurrence. Thirty-two patients who were diagnosed with SNIP and underwent surgery between 2010 and 2019 were enrolled: 24 men and 8 women, with a mean age of 59.2 years. The mean follow-up was 57.3 months. Demographics and information about history of smoking, diabetes mellitus (DM), hypertension, allergic rhinitis, alcohol consumption, tumor stage, surgical approach, and recurrence were reviewed retrospectively. Specimens were investigated using polymerase chain reaction to detect HPV DNA (high-risk subtypes: 16, 18, 31, 33, 35, 52b, and 58; low-risk subtypes: 6 and 11). Seven patients (21.9%) experienced recurrence. HPV DNA was detected in five (15.6%) patients (high-risk subtypes, n = 2; low-risk subtypes, n = 3). Patients with recurrence of SNIP had a higher proportion of young adults and displayed higher rates of HPV infection, DM, and advanced tumor stage than those without recurrence. HPV infection, young adulthood, DM, and advanced tumor stage could be associated with a high recurrence rate, which suggests that patients with these risk factors could require close follow-up after surgery

〔研究ノート〕 グリーンナッツオイルの摂取は マウスのDHAおよびEPAを増加させるか

In general, it has been recognized that n-3 fatty acids have the effect of suppressing inflammation and deterioration of cognitive ability. In addition, it is reported that a-linolenic acid is partially converted into DHA and EPA in the human body. On the other hand, like linseed oil and perilla oil, green nut oil （GNO） is rich in a-linolenic acid, one of the n-3 fatty acids, but not enough research on green nut oil has been done yet.Therefore, using mice, we investigated the effects of green nut oil intake on fatty acid composition in the liver, erythrocyte membrane, and brain. The result suggested that a-linolenic acid in green nut oil increases DHA and EPA in the bodies of mice. While mice that ingested corn oil （as a contrast） showed a slight decrease in brain DHA in 20-week-old mice as compared with 8-week-old mice, mice that ingested green nut oil showed no significant difference between 8 and 20-week-olds

Experience of Mothers of Hikikomori Children during their High School and University Days : The Process of Mother’s Participation in a Hikikomori “Parents’ Group”

The goal of this study is to clarify the experience process up to joining a parents' group, for mothers of school refusal children who refused to go to school or university and ended up as hikikomori, refusing to go out at all. Another goal is to examine support for such mothers. Semistructured interviews were conducted with twelve mothers. By analyzing the interviews using the revised version of the grounded theory approach, five categories and twelve concepts were extracted. The central category was looking for a guidepost and this affected the other three categories. In (makeshift solution for child) mothers repeatedly tried to find some stopgap solution. In (anguish amid a sense of isolation) they struggled on alone, feeling isolated. In this process, mothers made the (decision to join a parents' group) because they wanted to meet someone who would understand, obtain useful information and have some idea of what the future might hold. Since the looking for a guidepost situation continued for some time before they made the (decision to join a parents' group) it is highly likely that these mothers' psychological state can be stabilized through an appropriate response, and that they will be able to deal with their child head on, and adopt an attitude which seeks to understand him. From the analysis results, it is suggested that “mother's acceptance” and “encouragement of supporter's understanding of hikikomori” are important elements in supporting responses to looking for a guidepost.本研究は、高校・大学時にひきこもった子どもの母親の、親の会に参加するまでの体験のプロ セスを明らかにし支援の検討を目的とした。母親12名を対象に半構造化面接を行い、面接内容を 修正版グラウンデッド・セオリー・アプローチを用いて分析した結果、５つのカテゴリーと12の 概念が抽出された。コアカテゴリーは≪探し求める道しるべ≫であり、その後の３つのカテゴ リーに影響を与えていた。〈子どもには対症療法〉でその場しのぎの対応を繰り返し〈孤独感の 中で苦悩〉し、わかる人とつながり有効な情報を得て先の展望を持ちたいという思いで〈親の会 への参加決断〉をしていた。≪探し求める道しるべ≫状態が継続していることから、ここへの適 切な対応で早期に母親の心理的安定を図ることができる可能性が高く、“母親の受容”、”支援 者のひきこもりに関する理解の促進”が重要な要素と考えられる