Development of 1,3a,6a-triazapentalene-labeled enterobactin as a fluorescence quenching sensor of iron ion

1,3a,6a-Triazapentalene (TAP)-labeled enterobactin was developed as an iron ion sensor. 3-Acetylated-TAP was successfully introduced to the catechol ring of enterobactin, a well-recognized siderophore secreted by various Gram-negative bacteria. The fluorescence of TAPlabeled enterobactin decreased gradually as the amount of Fe3+ ion as an additive was increased, and 1.2 equiv of Fe3+ ion completely quenched the fluorescence. In clear contrast, when other metal ions were used, the fluorescence of TAP-labeled enterobactin remained even at 5.0 equiv

Periductal Induction of High Endothelial Venule-Like Vessels in Type 1 Autoimmune Pancreatitis

信州大学博士（医学）・学位論文・平成24年3月31日授与（甲第946号）・丸山 雅史This is a non-final version of an article published in final form in PANCREAS. 42(1):53-59 (2013).Objectives: Type 1 autoimmune pancreatitis (AIP) is histologically characterized by dense lymphoplasmacytic infiltration and marked storiform fibrosis, manifestations associated with pancreatic ducts. Such periductal lymphocyte recruitment is thought to be elicited by dysregulation of mechanisms governing physiological lymphocyte homing. The present study was undertaken to determine whether vascular addressins including peripheral lymph node addressin and mucosal addressin cell adhesion molecule 1 (MAdCAM-1) play a role in type 1 AIP histogenesis. Methods: Tissue sections of type 1 AIP and tumor-associated non-AIP chronic pancreatitis, as well as normal pancreas, were subjected to immunohistochemical analysis using vascular addressin-related antibodies. Results: The number of periductal mouse endothelial cell antigen 79-positive high endothelial venule (HEV)-like vessels was increased in type 1 AIP relative to that seen in non-AIP chronic pancreatitis, whereas the number of MAdCAM-1-positive HEV-like vessels did not differ between the 2 conditions. Mouse endothelial cell antigen 79 antigens are expressed on duct-forming epithelial cells not only in pancreas but also in salivary glands, which often harbor extrapancreatic lesions in type 1 AIP. Conclusions: Type 1 AIP can be characterized by periductal induction of MECA-79-positive HEV-like vessels. MECA-79-positive 6-sulfo sialyl Lewis X-related carbohydrate antigens expressed on duct-forming epithelial cells could be associated with type 1 AIP pathogenesis.ArticlePANCREAS. 42(1):53-59 (2013)journal articl

Efficient production of infectious viruses requires enzymatic activity of Epstein-Barr virus protein kinase

AbstractThe Epstein-Barr virus (EBV) BGLF4 gene product is the only protein kinase encoded by the virus genome. In order to elucidate its physiological roles in viral productive replication, we here established a BGLF4-knockout mutant and a revertant virus. While the levels of viral DNA replication of the deficient mutant were equivalent to those of the wild-type and the revertant, virus production was significantly impaired. Expression of the BGLF4 protein in trans fully complemented the low yield of the mutant virus, while expression of a kinase-dead (K102I) form of the protein failed to restore the virus titer. These results demonstrate that BGLF4 plays a significant role in production of infectious viruses and that the kinase activity is crucial

Degradation of Phosphorylated p53 by Viral Protein-ECS E3 Ligase Complex

p53-signaling is modulated by viruses to establish a host cellular environment advantageous for their propagation. The Epstein-Barr virus (EBV) lytic program induces phosphorylation of p53, which prevents interaction with MDM2. Here, we show that induction of EBV lytic program leads to degradation of p53 via an ubiquitin-proteasome pathway independent of MDM2. The BZLF1 protein directly functions as an adaptor component of the ECS (Elongin B/C-Cul2/5-SOCS-box protein) ubiquitin ligase complex targeting p53 for degradation. Intringuingly, C-terminal phosphorylation of p53 resulting from activated DNA damage response by viral lytic replication enhances its binding to BZLF1 protein. Purified BZLF1 protein-associated ECS could be shown to catalyze ubiquitination of phospho-mimetic p53 more efficiently than the wild-type in vitro. The compensation of p53 at middle and late stages of the lytic infection inhibits viral DNA replication and production during lytic infection, suggesting that the degradation of p53 is required for efficient viral propagation. Taken together, these findings demonstrate a role for the BZLF1 protein-associated ECS ligase complex in regulation of p53 phosphorylated by activated DNA damage signaling during viral lytic infection

Synthesis of yellow and red fluorescent 1,3a,6a-triazapentalenes and the theoretical investigation of their optical properties

To expand the originally developed fluorescent 1,3a,6a-triazapentalenes as fluorescent labelling reagents, the fluorescence wavelength of the 1,3a,6a-triazapentalenes was extended to the red color region. Based on the noteworthy correlation of the fluorescence wavelength with the inductive effect of the 2-substituent, electron-deficient 2-(2-cyano-4-methoxycarbonylphenyl)-1,3a,6a-triazapentalene and 2-(2,6-dicyano-4-methoxycarbonylphenyl)-1,3a,6a-triazapentalene were synthesized. The former exhibited yellow fluorescence and the latter exhibited red fluorescence, and both compounds exhibited large Stokes shifts, and the 1,3a,6a-triazapentalene system enabled the same fluorescent chromophore to cover the entire region of visible wavelengths. The potential applications of the 1,3a,6a-triazapentalenes as fluorescent probes in the fields of the life sciences were investigated, and the 1,3a,6a-triazapentalene system was clearly proven to be useful as a fluorescent reagent for live cell imaging. Quantum chemical calculations were performed to investigate the optical properties of the 1,3a,6a-triazapentalenes. These calculations revealed that the excitation involves a significant charge-transfer from the 1,3a,6a-triazapentalene skeleton to the 2-substituent. The calculated absorption and fluorescence wavelengths showed a good correlation with the experimental ones, and thus the system could enable the theoretical design of substituents with the desired optical properties

Expression of enhancer of zeste homolog 2 correlates with survival outcome in patients with metastatic breast cancer: exploratory study using primary and paired metastatic lesions

Evaluation of immunostaining of EZH2 and Ki67. (DOC 52 kb

Functionally confirmed compound heterozygous ADAM17 missense loss-of-function variants cause neonatal inflammatory skin and bowel disease 1

A disintegrin and metalloprotease 17 (ADAM17) is the major sheddase that processes more than 80 substrates, including tumour necrosis factor-α (TNFα). The homozygous genetic deficiency of ADAM17 causing a complete loss of ADAM17 expression was reported to be linked to neonatal inflammatory skin and bowel disease 1 (NISBD1). Here we report for the first time, a family with NISBD1 caused by functionally confirmed compound heterozygous missense variants of ADAM17, namely c.1699T>C (p.Cys567Arg) and c.1799G>A (p.Cys600Tyr). Both variants were detected in two siblings with clinical features of NISBD1, such as erythroderma with exudate in whole body, recurrent skin infection and sepsis and prolonged diarrhoea. In a cell-based assay using Adam10/17 double-knockout mouse embryonic fibroblasts (Adam10/17−/− mEFs) exogenously expressing each of these mutants, phorbol 12-myristate 13-acetate-stimulated shedding was strongly reduced compared with wild-type ADAM17. Thus, in vitro functional assays demonstrated that both missense variants cause the loss-of-function of ADAM17, resulting in the development of NISBD1. Our study further expands the spectrum of genetic pathology underlying ADAM17 in NISBD1 and establishes functional assay systems for its missense variants

障害受容における患者の意識変化と家族関係の一考察 : 在宅への移行期にある高齢患者への支援

著者が訪問看護を行った高齢者への関わりの振り返りを行い,入院中から退院後の生活にかけて,高齢者への看護のあり方の課題を得ることを目的に考察を加えた。疾病による障害を経験した高齢者が,どのような生きがいの変化を体験しているのか,また,生きがいに影響している因子との関連性があるのかを明らかにした。その結果,患者は退職という社会的な役割変化を経験し始める時期に発症することで,身体的な障害を受けると同時に,社会と家庭の両方において役割喪失を体験し,発症前の自己像との違いに葛藤している状態であった。しかし,家族から患者の気持ちを受け止めている発言がみられたことで,患者の表情や言葉に変化が見られ,障害受容につながっていると思われた。よって,障害を有する患者本人のありのまま全てを受け入れるか否かが,家族にとっての障害受容ではないかと考えられる。そして,それを受容した家族から受けるサポートによって,患者は障害受容ができると考えられる。このことから,専門職は,患者本人の障害受容と家族の受容の,2者に対しての支援を考えることが必要であることが示唆された。The purpose of this discussion was to reflect on our home-visit nursing program and the involvement of elderly patients, and clarify tasks for appropriate nursing of the elderly from the hospitalization period to the period after discharge. We evaluated changes in something to live for in elderly patients with disability due to disease, and factors affecting what they lived for. As a result, since patients developed disease near the retirement age when changes in their social role occur, they not only developed physical disability but also experienced the loss of roles both in society and at home, and experienced a conflict between their own images before and after the onset of the disease. However, after their families verbally expressed their acceptance of the patients\u27 feelings, the patients\u27 facial expressions and words changed, leading to their acceptance of the disability. Whether families accept all aspects of disabled patients is important. Support by families who have accepted patients may consequently enable patients to accept their own disabilities. Therefore, specialists should provide support to both patients and their families to facilitate this