Activation and contraction of human ‘vascular’ smooth muscle cells grown from circulating blood progenitors

Blood outgrowth smooth muscle cells offer the means to study vascular cells without the requirement for surgery providing opportunities for drug discovery, tissue engineering and personalised medicine. However, little is known about these cells which has meant their therapeutic potential remains unexplored. Our objective was to investigate for the first time the ability of blood outgrowth smooth muscle cells and vessel derived smooth muscle cells to sense the thromboxane mimetic U46619 by measuring intracellular calcium elevation and contraction. U46619 (10 26 -6 M) increased cytosolic calcium in blood outgrowth smooth muscle cells fibroblasts. Increased calcium signal peaked between 10-20 seconds after U46619 in both smooth muscle cell types. Importantly, U46619 (10-9 to 10-6 M) induced concentration-dependent contractions of both blood outgrowth smooth muscle cells and vascular smooth muscle cells but not in fibroblasts. In summary, we show that functional responses of blood outgrowth smooth muscle cells are in line with vascular smooth muscle cells providing critical evidence of their application in biomedical research

Required Elements in tRNA for Methylation by the Eukaryotic tRNA (Guanine-N2-) Methyltransferase (Trm11-Trm112 Complex)

The Saccharomyces cerevisiae Trm11 and Trm112 complex (Trm11-Trm112) methylates the 2-amino group of guanosine at position 10 in tRNA and forms N2-methylguanosine. To determine the elements required in tRNA for methylation by Trm11-Trm112, we prepared 60 tRNA transcript variants and tested them for methylation by Trm11-Trm112. The results show that the precursor tRNA is not a substrate for Trm11-Trm112. Furthermore, the CCA terminus is essential for methylation by Trm11-Trm112, and Trm11-Trm112 also only methylates tRNAs with a regular-size variable region. In addition, the G10-C25 base pair is required for methylation by Trm11-Trm112. The data also demonstrated that Trm11-Trm112 recognizes the anticodon-loop and that U38 in tRNAAla acts negatively in terms of methylation. Likewise, the U32-A38 base pair in tRNACys negatively affects methylation. The only exception in our in vitro study was tRNAValAAC1. Our experiments showed that the tRNAValAAC1 transcript was slowly methylated by Trm11-Trm112. However, position 10 in this tRNA was reported to be unmodified G. We purified tRNAValAAC1 from wild-type and trm11 gene deletion strains and confirmed that a portion of tRNAValAAC1 is methylated by Trm11-Trm112 in S. cerevisiae. Thus, our study explains the m2G10 modification pattern of all S. cerevisiae class I tRNAs and elucidates the Trm11-Trm112 binding sites

アンカースクリューを用いた上下顎前突治療例の長期保定

今回，我々は歯科矯正用アンカースクリューを用いて上下顎歯列全体の遠心移動を行うことで前歯部叢生および上下顎前歯の唇側傾斜を改善し，保定後3年経過時においても安定した咬合関係が維持された症例を経験したので報告する．患者は初診時年齢21歳7か月の女性で，上顎前突および叢生を主訴として来院した．側貌はconvex typeであり，上下口唇の突出と上下顎前歯部に軽度の叢生が認められた．大臼歯関係は両側Angle II級であり，オーバージェットは ＋3.0 mm，オーバーバイトは ＋1.4 mmであった．側面頭部エックス線規格写真より，∠ANBは5.1°とやや大きくskeletal Class IIを呈し，FMAは34.2°とhigh mandibular plane angle caseであった．歯系では，上顎中切歯歯軸傾斜角は標準的な値を示したが，下顎中切歯歯軸傾斜角は唇側傾斜を呈していた．U1 to NA は8.2 mm，L1 to NBは13.0 mmであり，上下顎中切歯はともに唇側に位置していた．治療方針として，上下顎前歯部の叢生および上下顎前歯唇側傾斜の解消のために，小臼歯抜去ではなく上下顎両側大臼歯部に歯科矯正用アンカースクリューを埋入し，上下顎歯列全体の遠心移動を行うこととした．2年9か月の動的治療の結果，前歯部叢生および口元の突出感は解消され，緊密な咬合が獲得された．保定後3年が経過した現在においても安定した咬合関係が保たれている．本症例より，歯科矯正用アンカースクリューを用いた上下顎歯列全体の遠心移動を行っても，長期間の安定が得られうることが示された

サンジゲン デジタル モデル オ モチイタ ヘンソクセイ コウシンレツ コウガイレツ カンジャ ノ コウゴウ ヒョウカ ニ カンスル ケントウ

【目的】Goslon Yardstickは片側性口唇裂口蓋裂患者の咬合状態から治療の難易度を評価する方法で，簡便かつ再現性が高いことから広く用いられている。しかし，評価に際し相当数の歯列模型を用いるため，その管理がしばしば困難となる。一方，三次元デジタルモデル（以下，3DDモデル）は石膏模型をデジタルデータに変換するため保存が容易であるが，デジタルデータより構築した画像は石膏模型とは異なった印象を受けることもある。今回我々は，石膏模型より作成した3DDモデルを用いてGoslon Yardstickによる評価を行い，口腔模型を用いた場合との差異について検討を行った。 【資料および方法】資料として，徳島大学病院矯正歯科を受診した片側性口唇裂口蓋裂患者37症例の石膏模型を用いた。それぞれの石膏模型をスキャニングし，3DDモデル（OrthoCAD，CADENT）を作成した。8年以上の臨床経験をもつ矯正歯科医4名が，石膏模型と3DDモデルをそれぞれ2回ずつ，5日以上の間隔をあけて評価を行った。評価はMarsらの方法に従った。また，評価の一致度を求めるために重み付きkappa値を算出した。kappa値は0.81～1.0をgood agreement，0.61～0.80をsubstantial，0.41～0.60をmoderate，0.21～0.40をfair，0.20以下をpoorとした。 【結果】石膏模型と3DDモデルについて評価者内でのkappa値はそれぞれ0.82～0.91，0.77～0.85となり，ともに評価は一致していた。このことから，石膏模型，3DDモデルのいずれを用いた場合でも評価の再現性が高いことが示された。さらに同一評価者内の石膏模型と3DDモデルの評価の一致度については，kappa値は0.75～0.86となり，石膏模型と3DDモデルにおける評価は比較的一致していた。一致していなかったものに着目すると，3DDモデルの方がGoslon Yardstickの値をやや高く評価する傾向にあった。 【考察】Goslon Yardstickの評価にあたって，3DDモデルの使用は石膏模型と比較しても遜色がないことが示された。三次元デジタルモデルを用いた片側性口唇裂口蓋裂患者の咬合評価に関する検

Intratracheal trimerized nanobody cocktail administration suppresses weight loss and prolongs survival of SARS-CoV-2 infected mice

新型コロナウイルスを中和するアルパカ抗体 --マウス実験で有効性を確認--. 京都大学プレスリリース. 2023-02-17.BACKGROUND: SARS-CoV-2 Omicron variants are highly resistant to vaccine-induced immunity and human monoclonal antibodies. METHODS: We previously reported that two nanobodies, P17 and P86, potently neutralize SARS-CoV-2 VOCs. In this study, we modified these nanobodies into trimers, called TP17 and TP86 and tested their neutralization activities against Omicron BA.1 and subvariant BA.2 using pseudovirus assays. Next, we used TP17 and TP86 nanobody cocktail to treat ACE2 transgenic mice infected with lethal dose of SARS-CoV-2 strains, original, Delta and Omicron BA.1. RESULTS: Here, we demonstrate that a novel nanobody TP86 potently neutralizes both BA.1 and BA.2 Omicron variants, and that the TP17 and TP86 nanobody cocktail broadly neutralizes in vitro all VOCs as well as original strain. Furthermore, intratracheal administration of this nanobody cocktail suppresses weight loss and prolongs survival of human ACE2 transgenic mice infected with SARS-CoV-2 strains, original, Delta and Omicron BA.1. CONCLUSIONS: Intratracheal trimerized nanobody cocktail administration suppresses weight loss and prolongs survival of SARS-CoV-2 infected mice

Spectral evolution of GRB 060904A observed with Swift and Suzaku -- Possibility of Inefficient Electron Acceleration

We observed an X-ray afterglow of GRB 060904A with the Swift and Suzaku satellites. We found rapid spectral softening during both the prompt tail phase and the decline phase of an X-ray flare in the BAT and XRT data. The observed spectra were fit by power-law photon indices which rapidly changed from $\Gamma = 1.51^{+0.04}_{-0.03}$ to $\Gamma = 5.30^{+0.69}_{-0.59}$ within a few hundred seconds in the prompt tail. This is one of the steepest X-ray spectra ever observed, making it quite difficult to explain by simple electron acceleration and synchrotron radiation. Then, we applied an alternative spectral fitting using a broken power-law with exponential cutoff (BPEC) model. It is valid to consider the situation that the cutoff energy is equivalent to the synchrotron frequency of the maximum energy electrons in their energy distribution. Since the spectral cutoff appears in the soft X-ray band, we conclude the electron acceleration has been inefficient in the internal shocks of GRB 060904A. These cutoff spectra suddenly disappeared at the transition time from the prompt tail phase to the shallow decay one. After that, typical afterglow spectra with the photon indices of 2.0 are continuously and preciously monitored by both XRT and Suzaku/XIS up to 1 day since the burst trigger time. We could successfully trace the temporal history of two characteristic break energies (peak energy and cutoff energy) and they show the time dependence of $\propto t^{-3} \sim t^{-4}$ while the following afterglow spectra are quite stable. This fact indicates that the emitting material of prompt tail is due to completely different dynamics from the shallow decay component. Therefore we conclude the emission sites of two distinct phenomena obviously differ from each other.Comment: 19 pages, 9 figures, accepted for publication in PASJ (Suzaku 2nd Special Issue