婦人科悪性腫瘍におけるTFの発現と過凝固状態についての研究

科学研究費助成事業 研究成果報告書：若手研究(B)2014-2017課題番号 : 2686131

Serotonergic Input to Orexin Neurons Plays a Role in Maintaining Wakefulness and REM Sleep Architecture

Neurons expressing neuropeptide orexins (hypocretins) in the lateral hypothalamus (LH) and serotonergic neurons in the dorsal raphe nucleus (DR) both play important roles in the regulation of sleep/wakefulness states, and show similar firing patterns across sleep/wakefulness states. Orexin neurons send excitatory projections to serotonergic neurons in the DR, which express both subtypes of orexin receptors (Mieda et al., 2011), while serotonin (5-HT) potently inhibits orexin neurons through activation of 5HT1A receptors (5HT1ARs). In this study, we examined the physiological importance of serotonergic inhibitory regulation of orexin neurons by studying the phenotypes of mice lacking the 5HT1A receptor gene (Htr1a) specifically in orexin neurons (ox5HT1ARKO mice). ox5HT1ARKO mice exhibited longer NREM sleep time along with decreased wakefulness time in the later phase of the dark period. We also found that restraint stress induced a larger impact on REM sleep architecture in ox5HT1ARKO mice than in controls, with a larger delayed increase in REM sleep amount as compared with that in controls, indicating abnormality of REM sleep homeostasis in the mutants. These results suggest that 5HT1ARs in orexin neurons are essential in the regulation of sleep/wakefulness states, and that serotonergic regulation of orexin neurons plays a crucial role in the appropriate control of orexinergic tone to maintain normal sleep/wake architecture

Long-Term Effect of Enzyme Replacement Therapy with Fabry Disease

Objective. To determine the effects of enzyme replacement therapy (ERT) on the hearing acuity in patients with Fabry disease. Materials. The study sample comprised 34 ears of 17 affected patients who underwent pure-tone audiometry before and after ERT. Methods. The patients were studied in relation to factors such as changes in hearing, presence of accompanying symptoms, status of renal and cardiac function, age, and gender. Data of pure-tone audiometry obtained before ERT and at the final examination were compared. Results. At the end of the follow-up period, no significant worsening of hearing acuity was noted at the end of the follow-up period. SSNHL was detected in 10 ears of 6 patients. Steroid therapy successfully cured the disease in 9 of the 10 ears. Conclusions. No significant worsening of hearing acuity was noted from the beginning to the end of ERT. The rate of improvement in SSNHL of Fabry disease was excellent in the treated patients. Hearing loss is a factor that causes marked deterioration of the patients’ quality of life, and it is desirable that the hearing acuity of patients be periodically evaluated and prompt treatment of SSNHL be administered, if available

Mechanism of the declining process of forest caused by the deposition of acid fog on forest canopies and the regeneration of the forest.

私達は、丹沢大山においてガス、エアロゾル、雨、霧、林内雨、樹幹流等の採取・分析を行った。関東平野に面するモミの樹冠には多量の汚染物質が沈着しているが、これはエーロゾルやガスの乾性沈着と霧の沈着の影響による。それぞれの寄与を見積もると、大山中腹ではガスの乾性沈着の影響が最も大きく、次いで霧水、粗大粒子、降雨の順であった。霧の発生頻度は標高の上昇と共に高くなるが、大気汚染物質の沈着量も同様であり、標高が高い地域の樹冠ほど霧を介した大気汚染物質の影響が大きい。丹沢山塊における微量ガス成分の濃度分布調査を行った結果、硝酸ガス等の濃度は大山より低いが大差無く、斜面方角依存性は小さいことが分かった。丹沢山塊では大気汚染物質の影響が大山とおよそ同レベルであり、その影響は標高の高い南東斜面の樹冠で顕著であり、ブナ衰退の斜面方角依存性には霧の影響が示唆された。大気汚染物質の植物影響を調べるため、ブナ苗木へのpH3の酸性霧曝露実験を行ったところ、葉の成長が悪くなり、落葉時期が早まった。酸性霧による同様な影響はモミについても見られるが、モミについての衰退メカニズムとしてまずワックス層が破壊され、続いて細胞壁を構成する糖鎖の架橋の機能を持つ2価のカルシウムイオンが水素イオンとイオン交換してはずれ、架橋が解けてカルシウムイオン、ホウ酸イオン、糖鎖が溶脱する。ブナについては蛍光顕微鏡を使って膜に結合したカルシウム濃度を調べたところ、酸の曝露により明らかに濃度減少することが示され、これによる耐性の減少が示唆された。以上のことから、大気汚染物質によりブナやモミの成長は抑制され、さらには枯れる可能性が高いこと、大気汚染物質の影響は大気汚染物質濃度だけでなく、霧の発生頻度、風向等に左右されるので植生の地形的環境に大きく依存することが明らかになった。In the mountain slope of Mt.Oyama, fog is frequently formed and pH of fogwater is low for nitric acid formed from NOx. The concentration of fog components becomes high when LWC is low, and acid fog events with fog water pH lower than 3.5 occurs for about 20% of fog duration there every year. It has been observed that the throughfall amount increased with the altitude for the contribution of fog, and the fog deposition may be dominated by the fog formation frequency and the deposition rate of fog droplets depending on the wind velocity. Deposited hydrogen ion via fog droplets is exchanged with metal ions in the cells in the leaves, and the concentrations of metal ions, calcium, magnesium, and potassium ions, in the throughfall is large for the leaching. In order to clarify the influence of acid fog on fir and beech, their seedlings were treated with simulated acid fog (SAF) at pH 3 and pH 5 prepared with a mixed solution of nitric acid, ammonium sulfate, sodium chloride with a molar ratio of 2:1:1 in a mobile fog chamber. The growth of the seedlings treated with the SAF at pH 3 had been reduced in plant height, stem diameter, the number of leaves, and dry matter production. Furthermore, the exposure to the SAF at pH 3 during growth phase induced early falling of leaves, and the falling rate were nearly 2 times greater than that at control. The laboratory experiment of the application of the simulated acid fog on their leaves also showed a large leaching of metal ions, and the leaching amount increased with the decreasing pH of the simulated fog solution. Not only the field observation but also laboratory experiments show the large effect of acid depositions on the trees.平成15年度～平成18年度科学研究費補助金（基盤研究（A））（課題番号：15201008

Sequencing analysis of 20,000 full-length cDNA clones from cassava reveals lineage specific expansions in gene families related to stress response

<p>Abstract</p> <p>Background</p> <p>Cassava, an allotetraploid known for its remarkable tolerance to abiotic stresses is an important source of energy for humans and animals and a raw material for many industrial processes. A full-length cDNA library of cassava plants under normal, heat, drought, aluminum and post harvest physiological deterioration conditions was built; 19968 clones were sequence-characterized using expressed sequence tags (ESTs).</p> <p>Results</p> <p>The ESTs were assembled into 6355 contigs and 9026 singletons that were further grouped into 10577 scaffolds; we found 4621 new cassava sequences and 1521 sequences with no significant similarity to plant protein databases. Transcripts of 7796 distinct genes were captured and we were able to assign a functional classification to 78% of them while finding more than half of the enzymes annotated in metabolic pathways in Arabidopsis. The annotation of sequences that were not paired to transcripts of other species included many stress-related functional categories showing that our library is enriched with stress-induced genes. Finally, we detected 230 putative gene duplications that include key enzymes in reactive oxygen species signaling pathways and could play a role in cassava stress response features.</p> <p>Conclusion</p> <p>The cassava full-length cDNA library here presented contains transcripts of genes involved in stress response as well as genes important for different areas of cassava research. This library will be an important resource for gene discovery, characterization and cloning; in the near future it will aid the annotation of the cassava genome.</p

Sequencing analysis of 20,000 full-length cDNA clones from cassava reveals lineage specific expansions in gene families related to stress response

<p>Abstract</p> <p>Background</p> <p>Cassava, an allotetraploid known for its remarkable tolerance to abiotic stresses is an important source of energy for humans and animals and a raw material for many industrial processes. A full-length cDNA library of cassava plants under normal, heat, drought, aluminum and post harvest physiological deterioration conditions was built; 19968 clones were sequence-characterized using expressed sequence tags (ESTs).</p> <p>Results</p> <p>The ESTs were assembled into 6355 contigs and 9026 singletons that were further grouped into 10577 scaffolds; we found 4621 new cassava sequences and 1521 sequences with no significant similarity to plant protein databases. Transcripts of 7796 distinct genes were captured and we were able to assign a functional classification to 78% of them while finding more than half of the enzymes annotated in metabolic pathways in Arabidopsis. The annotation of sequences that were not paired to transcripts of other species included many stress-related functional categories showing that our library is enriched with stress-induced genes. Finally, we detected 230 putative gene duplications that include key enzymes in reactive oxygen species signaling pathways and could play a role in cassava stress response features.</p> <p>Conclusion</p> <p>The cassava full-length cDNA library here presented contains transcripts of genes involved in stress response as well as genes important for different areas of cassava research. This library will be an important resource for gene discovery, characterization and cloning; in the near future it will aid the annotation of the cassava genome.</p

Oral hypofunction in the older population : Position paper of the Japanese Society of Gerodontology in 2016

Background: There is growing international interest in identifying the effects of ageing on oral health and on appropriate strategies for managing oral disorders. The Japanese Society of Gerodontology (JSG), as the official representative of researchers and clinicians interested in geriatric dentistry in Japan, makes several recommendations on the concept of “oral hypofunction.” Aims: This study proposes diagnostic criteria and management strategies to reduce the risk of oral hypofunction among older people. Conceptual Framework: We define oral hypofunction as a presentation of 7 oral signs or symptoms: oral uncleanness; oral dryness; decline in occlusal force; decline in motor function of tongue and lips; decline in tongue pressure; decline in chewing function; and decline in swallowing function. The criteria of each symptom were determined based on the data of previous studies, and oral hypofunction was diagnosed if the criteria for 3 or more signs or symptoms were met. Conclusions: We recommend that more evidence should be gathered from clinical studies and trials to clarify our diagnostic criteria and management strategies

Role of Sphingomyelinase in Infectious Diseases Caused by Bacillus cereus

Bacillus cereus (B. cereus) is a pathogen in opportunistic infections. Here we show that Bacillus cereus sphingomyelinase (Bc-SMase) is a virulence factor for septicemia. Clinical isolates produced large amounts of Bc-SMase, grew in vivo, and caused death among mice, but ATCC strains isolated from soil did not. A transformant of the ATCC strain carrying a recombinant plasmid containing the Bc-SMase gene grew in vivo, but that with the gene for E53A, which has little enzymatic activity, did not. Administration of an anti-Bc-SMase antibody and immunization against Bc-SMase prevented death caused by the clinical isolates, showing that Bc-SMase plays an important role in the diseases caused by B. cereus. Treatment of mouse macrophages with Bc-SMase resulted in a reduction in the generation of H2O2 and phagocytosis of macrophages induced by peptidoglycan (PGN), but no effect on the release of TNF-α and little release of LDH under our experimental conditions. Confocal laser microscopy showed that the treatment of mouse macrophages with Bc-SMase resulted in the formation of ceramide-rich domains. A photobleaching analysis suggested that the cells treated with Bc-SMase exhibited a reduction in membrane fluidity. The results suggest that Bc-SMase is essential for the hydrolysis of SM in membranes, leading to a reduction in phagocytosis

GABAergic neurons in the preoptic area send direct inhibitory projections to orexin neurons

Populations of neurons in the hypothalamic preoptic area (POA) fire rapidly during sleep, exhibiting sleep/waking state-dependent firing patterns that are the reciprocal of those observed in the arousal system. The majority of these preoptic "sleep-active" neurons contain the inhibitory neurotransmitter GABA. On the other hand, a population of neurons in the lateral hypothalamic area (LHA) contains orexins, which play an important role in the maintenance of wakefulness, and exhibit an excitatory influence on arousal-related neurons. It is important to know the anatomical and functional interactions between the POA sleep-active neurons and orexin neurons, both of which play important, but opposite roles in regulation of sleep/wakefulness states. In this study, we confirmed that specific pharmacogenetic stimulation of GABAergic neurons in the POA leads to an increase in the amount of non-rapid eye movement (NREM) sleep. We next examined direct connectivity between POA GABAergic neurons and orexin neurons using channelrhodopsin 2 (ChR2) as an anterograde tracer as well as an optogenetic tool. We expressed ChR2-eYFP selectively in GABAergic neurons in the POA by AAV-mediated gene transfer, and examined the projection sites of ChR2-eYFP-expressing axons, and the effect of optogenetic stimulation of ChR2-eYFP on the activity of orexin neurons. We found that these neurons send widespread projections to wakefulness-related areas in the hypothalamus and brain stem, including the LHA where these fibers make close appositions to orexin neurons. Optogenetic stimulation of these fibers resulted in rapid inhibition of orexin neurons. These observations suggest direct connectivity between POA GABAergic neurons and orexin neurons. © 2013 Saito, Tsujino, Hasegawa, Akashi, Abe, Mieda, Sakimura and Sakurai