Additional file 1: of Reporting quality of randomized controlled trials in patients with HIV on antiretroviral therapy: a systematic review

PRISMA checklist. (DOC 66 kb

Abrogation of PI3K signaling inhibits TGF-β1-mediated osteoblast differentiation in MC3T3-E1 cells.

<p>(A) Confluent MC3T3-E1 cells were cultured in OBM with and without 0.1 ng/mL TGF-β1 in the absence or presence of the PI3K inhibitor LY294002 (5, 10, or 25 µM) for 72 h. ALP activity was visualized by ALP staining of cells. (B) Cells were cultured in OBM with and without 0.1 ng/mL TGF-β1 in the absence or presence of 10 µM LY294002 for 72 h, and ALP activity was measured. Each experiment was performed in triplicate, and the data represent the means ± S.E. (<i>n</i> = 3). The Bonferroni correction for multiple comparisons was applied. *, <i>p</i><0.001.</p

Overexpression of CA-Akt reverses the inhibition of ALP activity induced by repeated administration of TGF-β1.

<p>MC3T3-E1 cells were infected with CA-Akt vector or Mock vector. Phosphorylation of Akt is evident in CA-Akt cells cultured in α-MEM (A). Then cells were treated with or without repeated administration of 0.1 ng/mL TGF-β1 for 72 h in OBM. The levels of phosphorylated Akt in these cells were detected by western blot analysis (B). ALP staining (C) and ALP activity (D) were assessed following repeated TGF-β1 administration. Figures shown represent at least three independent experiments. Values represent mean ± S.E. (<i>n</i> = 4). Bonferroni correction for multiple comparisons was applied. *, <i>p</i><0.001.</p

Akt Activation is Required for TGF-β1-Induced Osteoblast Differentiation of MC3T3-E1 Pre-Osteoblasts

<div><p>Background</p><p>We have previously reported that repeated treatment of human periodontal ligament cells and murine pre-osteoblast MC3T3-E1 cells with transforming growth factor-beta 1 (TGF-β1) inhibited their osteoblastic differentiation because of decreased insulin-like growth factor-1 (IGF-1) secretion. We also found that IGF-1/PI3K signaling plays an important role in osteoblast differentiation induced by TGF-β1 treatment; however, the downstream signaling controlling this remains unknown. The aim of this current study is to investigate whether Akt activation is required for osteoblast differentiation.</p><p>Methodology/Principal Findings</p><p>MC3T3-E1 cells were cultured in osteoblast differentiation medium (OBM) with or without 0.1 ng/mL TGF-β1. OBM containing TGF-β1 was changed every 12 h to provide repeated TGF-β1 administration. MC3T3-E1 cells were infected with retroviral vectors expressing constitutively active (CA) or dominant-negative (DN)-Akt. Alkaline phosphatase (ALP) activity and osteoblastic marker mRNA levels were substantially decreased by repeated TGF-β1 treatment compared with a single TGF-β1 treatment. However, expression of CA-Akt restored ALP activity following TGF-β1 treatment. Surprisingly, ALP activity increased following multiple TGF-β1 treatments as the number of administrations of TGF-β1 increased. Activation of Akt significantly enhanced expression of osteocalcin, but TGF-β1 treatment inhibited this. Mineralization of MC3T3-E1 cells was markedly enhanced by CA-Akt expression under all medium conditions. Exogenous IGF-1 restored the down-regulation of osteoblast-related gene expression by repeated TGF-β1 administration. However, in cells expressing DN-Akt, these levels remained inhibited regardless of IGF-1 treatment. These findings indicate that Akt activation is required for the early phase of osteoblast differentiation of MC3T3-E1 cells induced by TGF-β1. However, Akt activation is insufficient to reverse the inhibitory effects of TGF-β1 in the late stages of osteoblast differentiation.</p><p>Conclusions</p><p>TGF-β1 could be an inducer or an inhibitor of osteoblastic differentiation of MC3T3-E1 cells depending on the state of Akt phosphorylation. Our results indicate that Akt is the molecular switch for TGF-β1-induced osteoblastic differentiation of MC3T3-E1 cells.</p></div

Ion Sensitive Transparent-Gate Transistor for Visible Cell Sensing

In this study, we developed an ion-sensitive transparent-gate transistor (IS-TGT) for visible cell sensing. The gate sensing surface of the IS-TGT is transparent in a solution because a transparent amorphous oxide semiconductor composed of amorphous In–Ga–Zn-oxide (a-IGZO) with a thin SiO₂ film gate that includes an indium tin oxide (ITO) film as the source and drain electrodes is utilized. The pH response of the IS-TGT was found to be about 56 mV/pH, indicating approximately Nernstian response. Moreover, the potential signals of the IS-TGT for sodium and potassium ions, which are usually included in biological environments, were evaluated. The optical and electrical properties of the IS-TGT enable cell functions to be monitored simultaneously with microscopic observation and electrical measurement. A platform based on the IS-TGT can be used as a simple and cost-effective plate-cell-sensing system based on thin-film fabrication technology in the research field of life science

Nonoptical Detection of Allergic Response with a Cell-Coupled Gate Field-Effect Transistor

In this study, we report the label-free and reliable detection of allergic response using a cell-coupled gate field-effect transistor (cell-based FET). Rat basophilic leukemia (RBL-2H3) cells were cultured as a signal transduction interface to induce allergic reaction on the gate oxide surface of the FET, because IgE antibodies, which bind to Fcε receptors at the RBL-2H3 cell membrane, are specifically cross-linked by allergens, resulting in the allergic response of RBL-2H3 cells. In fact, the surface potential at the FET gate decreased owing to secretions such as histamine from the IgE-bound RBL-2H3 cells, which reacted with the allergen. This is because histamine, as one of the candidate secretions, shows basicity, resulting in a change in pH around the cell/gate interface. That is, the RBL-2H3-cell-based FET used in this study was originally from an ion-sensitive FET (ISFET), whose oxide surface (Ta₂O₅) with hydroxyl groups is fully responsive to pH on the basis of the equilibrium reaction. The allergic response of RBL-2H3 cells on the gate was also confirmed by estimating the amount of β-hexosaminidase released together with histamine and was analyzed using the electrical properties based on an inflammatory response of secreted histamine with the vascular endothelial cell-based FET. Thus, the allergic responses were monitored in a nonoptical and real-time manner using the cell-based FETs with the cellular layers on the gate, which reproduced the <i>in vivo</i> system and were useful for the reliable detection of the allergic reaction

Primers used for qRT-PCR.

<p>Primers used for qRT-PCR.</p

Forced expression of dominant-negative Akt abrogates the pro-differentiation effect of IGF-1.

<p>MC3T3-E1 cells were infected with DN-Akt vector or Mock vector (A), and then cells were cultured in OBM with and without a triple administration of 0.1 ng/mL TGF-β1 for 72 h. Exogenous IGF-1 (200 ng/mL) was applied in conjunction with administration of TGF-β1 and the expression of <i>Alp</i> and <i>Oc</i> mRNA levels measured in Mock (B, C) and DN-Akt (D, E) cells. Gene expression was analyzed by qRT-PCR, and mRNA levels were normalized to that of 18S rRNA and measured in triplicate. Values represent mean ± S.E. (<i>n</i> = 3). Bonferroni correction for multiple comparisons was applied. *, <i>p</i><0.001; ***, <i>p</i><0.05.</p

Repeated TGF-β1 treatment inhibits expression of osteoblast differentiation markers in MC3T3-E1 cells.

<p>Confluent MC3T3-E1 cells were treated for 72 h with OBM, OBM with a single administration of 0.1 ng/mL TGF-β1, and OBM with a triple administration of 0.1 ng/mL TGF-β1. Repeated TGF-β1 treatment significantly decreased the expression of <i>Alp</i> (A), <i>Oc</i> (C), and <i>Bsp</i> (D). The decrease in the expression of <i>Igf-1</i> (B) was not statistically significant. Expression of these genes was analyzed by qRT-PCR, and their mRNA levels were normalized to that of 18S rRNA and measured in triplicate. Values represent mean ± S.E. (<i>n</i> = 3). Bonferroni correction for multiple comparisons was applied. *, <i>p</i><0.001; **, <i>p</i><0.01; ***, <i>p</i><0.05.</p

The structure proposed for apteniol D is different from that of the compound obtained by total synthesis

<p>We describe the synthesis of 4,4′-oxyneolignan, the proposed structure for naturally occurring apteniol D. The diphenyl ether moiety in 4,4′-oxyneolignan was formed via classical Ullmann ether synthesis using excess copper powder in <i>N,N</i>-dimethylacetamide. The spectral data of synthesised apteniol D show differences compared to those of naturally occurring apteniol D.</p