Search

Cell extrusion is a universal mode of cell removal from tissues, and it plays an important role in regulating cell numbers and eliminating unwanted cells, such as apoptotic, unfit, or cancerous. During this process, cells delaminate from the cell layer, however, the underlying mechanisms remain to be elucidated. Here, we report a conserved execution mechanism of cell extrusion. We found extracellular vesicle (EV) formation in extruding cells at a site opposite to the extrusion direction. Particularly, we found that a lipid-scramblase‒mediated local exposure of phosphatidylserine is responsible for EV formation and is crucial for executing cell extrusion, while inhibition of this process disrupted prompt cell delamination and tissue homeostasis. Furthermore, we revealed that the EV formation is governed by the mechanism in microvesicle formation, while the EVs have some hallmarks of apoptotic body. Finally, we illustrated the role of EV formation as promoting the neighboring cells’ invasion resulting in the execution of cell extrusion by experimental and mathematical modeling analysis. Taken together, this study provides the insights that membrane dynamics plays a crucial role behind the cell exit from the tissue by connecting the actions of extruding cell and the neighboring cells.

When the four volumes of the Book of the Way of Incense (Kōdōsho) in the collection of the Mito City Museum were shown to the public for the first time in 1992, Akiyama Takashi contributed the introductory essay for the occasion. In this essay, Akiyama expressed doubts on the theory endorsed so far by Nishiyama Matsunosuke in his Iemoto seido no kenkyū（Research on the iemoto system）published in 1982 by Yoshikawa Kōbunkan, although he fell short of producing evidence against it. Nishiyama had stated that inside the domain of Mito, approximately the same territory as today’s Ibaraki prefecture, in regard to new disciples for the Shino School of incense “not even one was to be found for the entire period [of the Mito feudal domain]. It is thus conceivable that the way of incense was not accepted by the milieu in the Mito domain that developed Mitogaku”.In our research, drafted by three authors, we thoroughly examined the four volumes of the Book of the Way of Incense, consisting of Mito-shi hakuzō Kōnoki-jo (Introduction to Ko-awase, an incense matching game- Mito City Museum), Mito-shi hakuzō Kotokumikō (The earlier ten kinds of kumikō - Mito City Museum), Mito-shi hakuzō Hachikumikō (The eight kinds of kumikō- Mito City Museum) and Kōdō meikan (Compendium of the Way of Incense). While taking into account the previous studies by Midorikawa Fumiko and others, we also considered the historical documents of the Mito domain, with the purpose of investigate the relations between the domain of Mito and the way of incense in the early modern era (1603-1867).On one hand, we proceeded to reprint and revise the four original volumes of the Mito City Museum, adding explanatory notes, a translation into modern Japanese and related essays. The book we are currently writing with these contents will be entitled Kinseishoki no Kō-bunka (The culture of the incense in the early modern period) and its publication is planned for March 2020. It intends to clarify the culture of the incense in Kyoto (cultural capital) and in Mito (province) in the early modern period, from the year 1603 to the year 1700.On the other hand, in the present paper titled The incense tradition in the Mito domain we have broadened the outlook to the whole early modern period and considered the above-mentioned theory by Nishiyama.Consequently, it came to light that the first daimyō (feudal lord) of the Mito domain Tokugawa Yorifusa was enthusiastic about the way of incense, wrote a book himself on the subject and presented it to the Emperor Go-Mizunoo. Furthermore, Tokugawa Mitsukuni, the second daimyō of the Mito domain, presented the Emperor Go-sai with the book Fusō-shūyōshū, while both the fourth daimyō Munetaka and the eight daimyō Narinobu showed interest in the art of incense. The analysis of the documents also revealed that two vassals of Yorifusa, Okajima Matayuki and Iio Biku were practicing and researching the way of incense, proving that the tradition of incense continued until the latter part of the Edo period.However, regarding the way of incense in the Mito domain there are still aspects needing further investigation, which prevent us to reach a full clarification of the matter. Therefore, we are hereby releasing our findings in the form of research notes.

A 81-year-old man, who had been diagnosed in multiple cerebral infarction and Alzheimer's disease, was followed up in his local clinic since 1997. He had been bedridden before admission, but could eat. He was admitted with severe aspiration pneumonia in December 1999. Since severe dementia and dysphagia were noted after admission, he was examined to find out whether or not he could swallow while the treatment of his pneumonia was conducted at the same time. The water swallowing test indicated a risk of aspiration, thus, percutaneous endoscopic gastrostomy was performed on January 26, 2000 after the completion of the treatment for pneumonia. Although the patient's condition was complicated by aspiration pneumonia, enteral feeding through the gastric fistula gradually became successful, and he was discharged in June 2000. His family physician followed him up by visiting at home to examine and observe his general physical condition including consciousness, vital signs, skin and respiration, while taking measures in cooperation with the local health care visiting nurse. The patient, thereafter, was repeatedly admitted and discharged because of exacerbation and remission of symptoms, including coughing, sputum and fever, probably caused by aspiration pneumonia. When he was admitted in December 2001, which was his sixth admission, since there were troubles with the infusion tube and frequent gastroesophageal reflux, the gastric fistula management was judged to be a great burden on the patient. In January 2002, the gastrostomy tube was removed and the patients, whose alimentation was managed using intra-venous hyperalimentation (IVH), was discharged. Besides periodic visits by his family physician, a 24-hour house visit system was introduced to control his IVH and deal with his family members' anxiety. His general condition, thereafter, has not markedly changed. The patient has continuously received medical treatment for 14 months after being discharged and his condition is stable.

A 76-year-old HTLV-I-positive male with acute cerebellar ataxia was suffering from dyspnea on exertion. Chest CT suggested interstitial pneumonitis. Methylprednisolone pulse therapy improved his symptoms and chest CT findings. Twelve months after discharge, when the prednisolone dose was tapered to 5 mg every other day, his lung lesion recurred. The lesion responded initially to steroid therapy. However, hypoxemia intractable to steroid pulse therapy developed and the patient died of respiratory failure. The autopsy revealed diffuse alveolar hemorrhage with no finding of vasculitis. This is the first case report of diffuse alveolar hemorrhage in an HTLV-I carrier.

Background: Recent reports indicate that in response to various stimuli, eosinophils produce a variety of cytokines (e.g. IL–8) which play pivotal roles in allergic inflammation. In that regard, the transcription factor, nuclear factor, Kappa B (NF–κB), is an important activator of tumor–necrosis–factor–alpha (TNF–α)–induced IL–8 gene expression in monocytes, lymphocytes and neutrophils. We therefore investigated the role played by NF–κB in cytokine production induced by stimulation of eosinophils with the proinflammatory cytokines, granulocyte–monocyte colony–stimulating factor (GM–CSF) and TNF–α. Methods: Peripheral blood samples were obtained from human subjects with slight to moderate eosinophilia. NF–κB activation elicited by exposing cells to GM–CSF and/or TNF–α was investigated using immunohistochemistry and gel shift assays. To functionally assess the effects of NF–κB translocation, IL–8 production was also examined using an enzyme–linked immunosorbent assay. Results: Stimulation of eosinophils with GM–CSF + TNF–α induced significant increases in the synthesis and secretion of IL–8 which were associated with translocation of NF–κB p50 into the nucleus. The binding of NF–κB to the DNA was verified by the gel shift assays. IL–8 production was significantly inhibited by N–acetyl–L–cysteine, FK506 and MG–132, inhibitors of NF–κB activation and translocation. Conclusion: On the basis of our findings, we conclude that activation and translocation of NF–κB plays a crucial role in the signal–transduction pathway leading to the synthesis and release of IL–8 by eosinophils.

The mechanism for incorporation of aphidicolin-sensitive DNA polymerase into reconstituting sperm nuclei was studied in a Xenopus egg extract cell-free system. Aphidicolin-sensitive DNA polymerase activity was sedimented along with the light membrane fraction of Xenopus egg extract on a discontinuous sucrose gradient. Treatment of the egg extract with Triton X-100 caused DNA polymerase activity to migrate to a lighter density position at which free proteins were distributed. DNA polymerase activity was incorporated into the reconstituting sperm nuclei from the egg extract, but no nuclear incorporation was observed in nuclei incubated in egg extracts which had been treated with Triton X-100 or sonicated. The incorporation was also prohibited by several different treatments of the egg extract resulting in incomplete assembly of the nuclear membrane on the sperm nuclei. On the other hand, there was no inhibition of nuclear incorporation into the sperm nuclei reconstituting in the extracts which had been depleted of WGA-binding pore complex proteins or which contained a specific inhibitor of topoisomerase II (ICRF-193). In these two cases, the nuclear double-layered membrane assembled normally, although in the former case the sperm nuclei lacked lamina and did not initiate DNA replication, and in the latter case the sperm nuclei did not decondense but initiated DNA replication. Thus, it is concluded that DNA polymerase activity is incorporated into the reconstituting nuclei via the membraneous/particulate fraction of the egg extract simultaneously with nuclear double-layered membrane assembly. The lamina assembly and the transport system via the nuclear envelope pore complex are suggested not to participate in DNA polymerase nuclear incorporation.

A 81-year-old man, who had been diagnosed in multiple cerebral infarction and Alzheimer's disease, was followed up in his local clinic since 1997. He had been bedridden before admission, but could eat. He was admitted with severe aspiration pneumonia in December 1999. Since severe dementia and dysphagia were noted after admission, he was examined to find out whether or not he could swallow while the treatment of his pneumonia was conducted at the same time. The water swallowing test indicated a risk of aspiration, thus, percutaneous endoscopic gastrostomy was performed on January 26, 2000 after the completion of the treatment for pneumonia. Although the patient's condition was complicated by aspiration pneumonia, enteral feeding through the gastric fistula gradually became successful, and he was discharged in June 2000. His family physician followed him up by visiting at home to examine and observe his general physical condition including consciousness, vital signs, skin and respiration, while taking measures in cooperation with the local health care visiting nurse. The patient, thereafter, was repeatedly admitted and discharged because of exacerbation and remission of symptoms, including coughing, sputum and fever, probably caused by aspiration pneumonia. When he was admitted in December 2001, which was his sixth admission, since there were troubles with the infusion tube and frequent gastroesophageal reflux, the gastric fistula management was judged to be a great burden on the patient. In January 2002, the gastrostomy tube was removed and the patients, whose alimentation was managed using intra-venous hyperalimentation (IVH), was discharged. Besides periodic visits by his family physician, a 24-hour house visit system was introduced to control his IVH and deal with his family members' anxiety. His general condition, thereafter, has not markedly changed. The patient has continuously received medical treatment for 14 months after being discharged and his condition is stable.

Two fungi were isolated from soil which grew on 0.1_??_0.2% formaldehyde as the sole carbon source, and identified as Gliocladium deliquescens and Paecilomyces varioti. Both the strains could grow on 5% methanol and 5% Na-formate, while the former could grow even on 7% methanol. Metabolic pathways were traced through two dimensional paper chromatography and autoradiographic techniques using 14C-formaldehyde, 14C-methanol or 14C-CO2 as substrates. The intracellular metabolites were persued and their quantitative variation with time was measured. Along with the fact that serine and malate appeared in the earlier time, then appeared organic acids and amino acids belonging to TCA cycle, and the fact that hydroxy-pyruvate reductase and phosphoenolpyruvate carboxylase activities were much stronger in methanol culture than in ethanol culture, it was concluded that the two fungi followed the serine pathway in assimilating C1-compounds. Oxidation enzymes of methanol and formaldehyde were also studied, and an oxidizing system was found besides usual NAD linked methanol or formaldehyde dehydrogenases.