Carrier cell-mediated cell lysis of squamous cell carcinoma by squamous cell carcinoma antigen 1 promoter-driven oncolytic adenovirus

The squamous cell carcinoma antigen (SCCA) serves as a serological marker for squamous cell carcinomas. Molecular cloning of the SCCA genomic region has revealed the presence of two tandemly arrayed genes, SCCA1 and SCCA2. We examined the promoter activity of the 5'-flanking proximal region of the SCCA1 gene. Deletion analysis of SCCA1 promoter identified a 175-bp core promoter region and an enhancer region at -525 to -475 bp upstream of the transcription start site. The transcriptional activity of the SCCA1 promoter was up-regulated in squamous cell carcinoma cells, compared with normal keratinocyte, normal non-keratinocyte and adenocarcinoma cells. Five tandem repeats of enhancer increased SCCA1 promoter activity by 4-fold. Oncolytic adenovirus driven by the SCCA1 promoter with 5 tandem repeats of enhancer specifically killed squamous cell carcinoma cells in vitro and in vivo. A549 carrier cells infected with the oncolytic adenovirus induced complete regression of tumor by overcoming immunogenicity and adenovirus-mGM-CSF augmented the antitumor effect of carrier cells. These findings suggest that SCCA1 promoter is a potential target of gene therapy for squamous cell carcinoma

Ward Chief Nurses' Perceptions and the Current Situation of Nurses' Career Development

目的：近畿，中国・四国地方の地域密着型病院の病棟看護師長のキャリアアップに対する認識と現状を明らかにする．方法：研究協力が得られた病棟看護師長187名に自記式質問紙調査を実施した．結果：101名（回収率54．0％）から回答が得られた．54名がスタッフに進学希望者がおり，それに対する思い・考えは【看護師個人の知識・スキルアップ・キャリアアップにつながる】【病棟・病院の看護の質向上が図れる】【病棟管理者としてスタッフの成長・キャリアアップを支援したい】の３つに分類された．支援はスタッフのキャリアプランの目標管理と学習への指導・助言，勤務調整・研修費などの体制支援を行っているが，スタッフのキャリアへの志向性が低い，人員不足・勤務業務調整が難しいなどの課題が明らかとなった．結論：病棟看護師長は，スタッフの成長と看護の質向上を期待し，キャリアアップへの課題に対する支援を模索していることが伺えた．Objective : To clarify the perceptions of ward chief nurses at community-based hospitals in the Kinki, Chugoku, and Shikoku regions, as well as the current situation regarding career development. Methods : We conducted a self-administered questionnaire survey of 187 ward chief nurses. Results : Responses were obtained from 101 chief nurses （response rate : 54.0%）. Of the respondents, 54 replied that their ward had staff members who desired more advanced education. Their thoughts/opinions about advanced education were classified as follows :［ it helps those nurses to advance their knowledge/skills/career］, ［it is helpful in improving the quality of nursing care in wards/hospitals］, and ［（they）, as ward managers, want to support staff members’ growth/career development］. Support provided to staff members included guidance/advice on goal management, learning for career development, and organizational support such as work shift adjustment and training fees. On the other hand, issues included staff members’ low career orientation, manpower shortages, and difficulties in work shift and duty adjustment. Conclusion : The ward chief nurses wanted to encourage staff members’ growth and qualitative improvements in nursing care and to explore support for career development issues

Alterations in erythrocyte membrane lipid and its fragility in a patient with familial lecithin : cholesterol acyltrasferase (LCAT) deficiency

Lecithin : cholesterol acyltrasferase (LCAT) plays a key role in the cholesterol metabolism-mediated esterification of free cholesterol into the cholesterol ester in normal plasma. Familial LCAT deficiency is frequently associated with anemia. Using biochemical and physiological techniques, the erythrocytes of this patient were investigated to gain an insight into the relationship between the abnormalities of lipid metabolism and erythrocyte membrane fragility. Abnormal erythrocytes, so-called Target cells and/or Knizocytes, were observed at 20% in our patient’s erythrocytes. Moreover, the mean corpuscular volume of the patient’s cells was 7% greater than that of a normal individual. In the membrane lipids of the patient’s erythrocytes, cholesterol and phosphatidylcholine increased, and phosphatidylethanolamine decreased. The electron spin resonance technique with a fatty acid spin probe showed that the membrane fluidity was more elevated than that of normal cells in spite of the increase in cholesterol content and the cholesterol/ phospholipid ratio of the membrane of patient’s erythrocytes. The patient’s abnormally shaped erythrocytes were less deformed than those of the normal individual under high shear stress. The partial depletion of membrane cholesterol from the patient’s erythrocytes was demonstrated by incubation with normal plasma with LCAT activity. The increment of transformed erythrocytes during the incubation could be prevented by cholesterol depletion from the patient’s erythrocyte membrane. These findings indicate that normochromic anemia of the patient might be caused by erythrocyte fragility resulting from decreased deformity and/or abnormal shape of the cells due to abnormal lipid composition in the membrane

Tetranins: new putative spider mite elicitors of host plant defense

Summary The two‐spotted spider mite (Tetranychus urticae) is a plant‐sucking arthropod herbivore that feeds on a wide array of cultivated plants. In contrast to the well‐characterized classical chewing herbivore salivary elicitors that promote plant defense responses, little is known about sucking herbivores' elicitors. To characterize the sucking herbivore elicitors, we explored putative salivary gland proteins of spider mites by using an Agrobacterium‐mediated transient expression system or protein infiltration in damaged bean leaves. Two candidate elicitors (designated as tetranin1 (Tet1) and tetranin2 (Tet2)) triggered early leaf responses (cytosolic calcium influx and membrane depolarization) and increased the transcript abundances of defense genes in the leaves, eventually resulting in reduced survivability of T. urticae on the host leaves as well as induction of indirect plant defenses by attracting predatory mites. Tet1 and/or Tet2 also induced jasmonate, salicylate and abscisic acid biosynthesis. Notably, Tet2‐induced signaling cascades were also activated via the generation of reactive oxygen species. The signaling cascades of these two structurally dissimilar elicitors are mostly overlapping but partially distinct and thus they would coordinate the direct and indirect defense responses in host plants under spider mite attack in both shared and distinct manners

Arabidopsis receptor-like protein30 and receptor-like kinase suppressor of BIR1-1/EVERSHED mediate innate immunity to necrotrophic fungi

Effective plant defense strategies rely in part on the perception of non-self determinants, so-called microbe-associated molecular patterns (MAMPs), by transmembrane pattern recognition receptors leading to MAMP-triggered immunity. Plant resistance against necrotrophic pathogens with a broad host range is complex and yet not well understood. Particularly, it is unclear if resistance to necrotrophs involves pattern recognition receptors. Here, we partially purified a novel proteinaceous elicitor called SCLEROTINIA CULTURE FILTRATE ELICITOR1 (SCFE1) from the necrotrophic fungal pathogen Sclerotinia sclerotiorum that induces typical MAMP-triggered immune responses in Arabidopsis thaliana. Analysis of natural genetic variation revealed five Arabidopsis accessions (Mt-0, Lov-1, Lov-5, Br-0, and Sq-1) that are fully insensitive to the SCFE1-containing fraction. We used a forward genetics approach and mapped the locus determining SCFE1 sensitivity to RECEPTOR-LIKE PROTEIN30 (RLP30). We also show that SCFE1-triggered immune responses engage a signaling pathway dependent on the regulatory receptor-like kinases BRASSINOSTEROID INSENSITIVE1-ASSOCIATED RECEPTOR KINASE1 (BAK1) and SUPPRESSOR OF BIR1-1/EVERSHED (SOBIR1/EVR). Mutants of RLP30, BAK1, and SOBIR1 are more susceptible to S. sclerotiorum and the related fungus Botrytis cinerea. The presence of an elicitor in S. sclerotiorum evoking MAMP-triggered immune responses and sensed by RLP30/SOBIR1/BAK1 demonstrates the relevance of MAMP-triggered immunity in resistance to necrotrophic fungi

Specific proteolytic cleavage of agrin regulates maturation of the neuromuscular junction

During the initial stage of neuromuscular junction (NMJ) formation, nerve-derived agrin cooperates with muscle-autonomous mechanisms in the organization and stabilization of a plaque-like postsynaptic specialization at the site of nerve-muscle contact. Subsequent NMJ maturation to the characteristic pretzel-like appearance requires extensive structural reorganization. We found that the progress of plaque-to-pretzel maturation is regulated by agrin. Excessive cleavage of agrin via transgenic overexpression of an agrin-cleaving protease, neurotrypsin, in motoneurons resulted in excessive reorganizational activity of the NMJs, leading to rapid dispersal of the synaptic specialization. By contrast, expression of cleavage-resistant agrin in motoneurons slowed down NMJ remodeling and delayed NMJ maturation. Neurotrypsin, which is the sole agrin-cleaving protease in the CNS, was excluded as the physiological agrin-cleaving protease at the NMJ, because NMJ maturation was normal in neurotrypsin-deficient mice. Together, our analyses characterize agrin cleavage at its proteolytic α- and β-sites by an as-yet-unspecified protease as a regulatory access for relieving the agrin-dependent constraint on endplate reorganization during NMJ maturation

Spatial constraints dictate glial territories at murine neuromuscular junctions

Spatial competition between glial cells causes them to partition neuromuscular junctions into discrete domains within a synapse