Cell-free synthesis of alkaline lipase, a glyoxysomal membrane protein, from castor bean endosperm

AbstractPolyadenylated RNA from castor bean endosperm was translated in a wheat germ cell-free protein synthesis system, and alkaline lipase, an integral glyoxysomal membrane protein, was immunoprecipitated. The apparent molecular mass of the lipase synthesized in vitro was slightly higher than that of the mature enzyme (62 kDa). When mRNA derived from free and membrane-bound polysomes in the endosperms was translated in vitro, the lipase was predominantly recovered from the products of the free polysomal mRNA, suggesting that the membrane protein is post-translationally inserted into the membrane. The amino-terminal 8 amino acid residues of the mature lipase were sequenced

TANK prevents IFN-dependent fatal diffuse alveolar hemorrhage by suppressing DNA-cGAS aggregation

Diffuse alveolar hemorrhage (DAH) is one of the serious complications associated with systemic lupus erythematosus, an autoimmune disease whose pathogenesis involves type I IFNs and cytokines. Here, we show that TANK, a negative regulator of the NF-κB signaling via suppression of TRAF6 ubiquitination, is critical for the amelioration of fatal DAH caused by lung vascular endothelial cell death in a mouse model of systemic lupus erythematosus. The development of fatal DAH in the absence of TANK is mediated by type I IFN signaling, but not IL-6. We further uncover that STING, an adaptor essential for the signaling of cytoplasmic DNA sensor cyclic GMP-AMP (cGAMP) synthase (cGAS), plays a critical role in DAH under Tank deficiency. TANK controls cGAS-mediated cGAMP production and suppresses DNA-mediated induction of IFN-stimulated genes in macrophages by inhibiting the formation of DNA-cGAS aggregates containing ubiquitin. Collectively, TANK inhibits the cGAS-dependent recognition of cytoplasmic DNA to prevent fatal DAH in the murine lupus model

Physiological roles of Runx3 in female reproductive organs in mice

Runx3(Runtdomaintranscriptionfactor3)はRunxファミリーに属する転写因子で雌マウスにおいてRunx3 mRNAは, 卵巣や子宮に発現していた。雌のRunx3(-/-)マウスは不妊であった。Runx3(-/-)マウスは卵胞形成異常を起こしており, 無排卵であった。一方で, 排卵能および黄体形成能は有していた。以上より, Runx3は卵胞形成および排卵制御に関与していることを明らかにした。Runx3(-/-)マウスの子宮は萎縮している。子宮内膜上皮細胞では, E2依存性の細胞増殖が起こらなかった。しかし,子宮内膜間質細胞では, E2, P4存在下で正常に細胞増殖が起きた。以上より, Runx3はE2による子宮の細胞増殖に関与していることを明らかにした

Microglia in Alzheimer's Disease: Risk Factors and Inflammation

Microglia are resident immune cells in the central nervous system (CNS) that originate from myeloid progenitor cells in the embryonic yolk sac and are maintained independently of circulating monocytes throughout life. In the healthy state, microglia are highly dynamic and control the environment by rapidly extending and retracting their processes. When the CNS is inflamed, microglia can give rise to macrophages, but the regulatory mechanisms underlying this process have not been fully elucidated. Recent genetic studies have suggested that microglial function is compromised in Alzheimer's disease (AD), and that environmental factors such as diet and brain injury also affect microglial activation. In addition, studies of triggering receptor expressed on myeloid cells 2-deficiency in AD mice revealed heterogeneous microglial reactions at different disease stages, complicating the therapeutic strategy for AD. In this paper, we describe the relationship between genetic and environmental risk factors and the roles of microglia in AD pathogenesis, based on studies performed in human patients and animal models. We also discuss the mechanisms of inflammasomes and neurotransmitters in microglia, which accelerate the development of amyloid-β and tau pathology

Synthesis of 13a-methylphenanthroindolizidines using radical cascade cyclization: synthetic studies toward (±)-hypoestestatin 1

金沢大学大学院自然科学研究科生理活性物質科学A radical cascade involving 6-endo cyclization of aryl radicals generated from N-acryloyl-N-(1-methylethenyl)-9-bromophenanthren-10-ylmethylamines, followed by 5-endo-trig cyclization of the resulting α-amidoyl radicals afforded phenanthroindolizidines bearing a methyl substituent at the angular C13a position. 2,3,6-Trimethoxy derivative was synthesized by using this method, but its spectral data were not in accord with those of literature values reported for hypoestestatin 1. Further synthetic study toward hypoestestatin 1 is demonstrated. © 2007 Elsevier Ltd. All rights reserved

Transient and permanent gene transfer into the brain of the teleost fish medaka (Oryzias latipes) using human adenovirus and the Cre-loxP system

AbstractIn this study, we demonstrated that human type-5 adenovirus infected the brain of the teleost fish, medaka (Oryzias latipes), in vivo. Injection of adenoviral vector into the mesencephalic ventricle of medaka larvae induced the expression of reporter genes in some parts of the telencephalon, the periventricular area of the mesencephalon and diencephalon, and the cerebellum. Additionally, the Cre-loxP system works in medaka brains using transgenic medaka carrying a vector containing DsRed2, flanked by loxP sites under control of the β-actin promoter and downstream promoterless enhanced green fluorescent protein (EGFP). We demonstrated that the presence of green fluorescence depended on injection of adenoviral vector expressing the Cre gene and confirmed that EGFP mRNA was transcribed in the virus-injected larvae

Two distinct prions in fatal familial insomnia and its sporadic form

Abstract Fatal familial insomnia is a genetic prion disease, which is associated with the aspartic acid to asparagine substitution at codon 178 of the prion protein gene. Although the hallmark pathological feature is thalamic and olivary degeneration, there is a patient with an atypical fatal familial insomnia without the hallmark feature. The cause of the pathological variability is unclear. We analysed a Japanese fatal familial insomnia kindred and compared one atypical clinicopathological fatal familial insomnia phenotype case and typical fatal familial insomnia phenotype cases with transmission studies using multiple lines of knock-in mice and with protein misfolding cyclic amplification. We also analysed the transmissibility and the amplification properties of sporadic fatal insomnia. Transmission studies revealed that the typical fatal familial insomnia with thalamic and olivary degeneration showed successful transmission only using knock-in mice expressing human–mouse chimeric prion protein gene. The atypical fatal familial insomnia with spongiform changes showed successful transmission only using knock-in mice expressing bank vole prion protein gene. Two sporadic fatal insomnia cases with thalamic and olivary degeneration showed the same transmissibility as the typical fatal familial insomnia phenotype. Interestingly, one sporadic fatal insomnia case with thalamic/olivary degeneration and spongiform changes showed transmissibility of both the typical and atypical fatal familial insomnia phenotypes. Protein misfolding cyclic amplification could amplify both typical fatal familial insomnia cases and sporadic fatal insomnia cases but not the atypical fatal familial insomnia phenotype or other sporadic Creutzfeldt–Jakob disease subtypes. In addition to clinical findings and neuropathological features, the transmission properties and the amplification properties were different between the typical and atypical fatal familial insomnia phenotypes. It is suggested that two distinct prions were associated with the diversity in the fatal familial insomnia phenotype, and these two prions could also be detected in sporadic fatal insomnia

ニホン シカ ダイガク リンショウ ジッシュウ シサツ ホウコク

We made a field trip to Nippon Dental University Hospital to observe its clinical training practices on February 16, 2011. Distinctive features of the clinical training system were as follows. 1) Clinical training is given to fifth-year students. Students in the sixth-year are intent on their studies. 2) There is a one-month overlap in the training periods between these two groups, as patients are handed over from the fifth- to the fourth-year students. This handover is a student-led event. 3) After this period, pre-clinical training is given to the fifth-year students for three months, and then clinical training begins. 4) Patients are requested to evaluate and give feedback to the students treating them. Each student should receive five or more evaluations. 5) A mentor system was introduced in 2005. According to our findings and the results of the“ World Café” held on the same day with trainee dentists and fifth-year students, we identified the following means of improving our clinical training system. First, students under clinical training are now permitted to participate in case-report conferences of trainee dentists, as of 2011. This program was introduced based on the concept of top-down processing to help students form perceptions about cases and treatment. Second, the necessity of a faculty-development program focused on clinical training is recognized, to standardize and improve the guidance given to students by advising doctors. Third, the mentor system and the“ World Café” need more attention as elements of clinical training to support students and motivate improvement