Two Cases of Severe Degeneration of the Macula Following Vitrectomy with Indocyanine Green-Assisted Internal Limiting Membrane Peeling for Idiopathic Macular Hole

We describe three eyes of two cases of severe degeneration of the macula following vitrectomy with indocyanine green-assisted internal limiting membrane peeling for idiopathic macular hole. We need to remember the possibility of these complications and have to select the procedures that are safest to use for macular hole surgery

A Versatile and Rapidly Deployable Device to Enable Spatiotemporal Observations of the Sessile Microbes and Environmental Surfaces

Although microbes typically associate with surfaces, detailed observations of surface-associated microbes on natural substrata are technically challenging. We herein introduce a flow channel device named the Stickable Flow Device, which is easily configurable and deployable on various surfaces for the microscopic imaging of environmental microbes. We demonstrated the utility of this device by creating a flow channel on different types of surfaces including live leaves. This device enables the real-time imaging of bacterial biofilms and their substrata. The Stickable Flow Device expands the limits of conventional real-time imaging systems, thereby contributing to a deeper understanding of microbe-surface interactions on various surfaces

Upregulation of Mir-21 Levels in the Vitreous Humor Is Associated with Development of Proliferative Vitreoretinal Disease.

MicroRNAs (miRNAs) are small noncoding RNAs that regulate gene expression by post-transcriptional inhibition of mRNA translation. Dysregulation of miRNAs, including circulating miRNAs, has been reported to play an important role in the development of various diseases, including fibrotic diseases. Aberrant expression of miRNAs in the vitreous humor of vitreoretinal diseased eyes has been reported. However, the expression pattern of miRNAs present in the vitreous humor of proliferative vitreoretinal disease (PVD) patients, including proliferative diabetic retinopathy (PDR), and proliferative vitreoretinopathy (PVR), remains unknown. To investigate the factors important for the development of PVD, we characterized the miRNAs present in the vitreous humor of PVD patients and analyzed the expression profiles of 377 miRNAs using quantitative polymerase chain reaction-based miRNA arrays. The expression of a specific subset of miRNAs, previously reported to be associated with the development of angiogenesis and fibrosis, was significantly altered in the vitreous of PVD patients. Among these miRNAs, we identified miR-21 as a candidate fibrotic miRNA with an important role in the pathogenesis of PVD. Increased miR-21 levels in the vitreous were associated with retinal fibrosis, including PVR and PDR. Because epithelial-mesenchymal transition (EMT) of retinal pigment epithelial cells (RPECs) plays a critical role in retinal fibrosis, the expression of miR-21 in human RPECs was determined. Its expression in RPECs was induced by transforming growth factor-β, a key growth factor involved in fibrogenesis, and was enhanced by high glucose culture conditions, suggesting that miR-21 expression positively correlates with disease progression. Gain- and loss-of-function studies revealed that miR-21 promoted cell proliferation and migration of ARPE-19 cells without affecting EMT-related gene expression. Together, our studies have identified miR-21 as a potential disease-modifying miRNA in the vitreous humor that is involved in the development of retinal fibrosis and may be a novel marker of PVD

Unique Biological Activity and Potential Role of Monomeric Laminin-γ2 as a Novel Biomarker for Hepatocellular Carcinoma: A Review

Laminin (Ln)-332 consists of α3, β3, and γ2 chains, which mediate epithelial cell adhesion to the basement membrane. Ln-γ2, a component of Ln-332, is frequently expressed as a monomer in the invasion front of several types of malignant tissues without simultaneous expression of Ln-α3 and/or Ln-β3 chains. Moreover, monomeric Ln-γ2 induces tumor cell proliferation and migration in vitro. These unique biological activities indicate that monomeric Ln-γ2 could be a candidate biomarker for early cancer surveillance. However, the present immune method for monomeric Ln-γ2 detection can only predict its expression, since no antibody that specifically reacts with monomeric γ2, but not with heterotrimeric γ2 chain, is commercially available. We have, therefore, developed monoclonal antibodies to specifically detect monomeric Ln-γ2, and devised a highly sensitive method to measure serum monomeric Ln-γ2 levels using a fully automated chemiluminescent immunoassay (CLIA). We evaluated its diagnostic value in sera from patients with several digestive cancers, including hepatocellular carcinoma (HCC), and found serum monomeric Ln-γ2 to be a clinically available biomarker for HCC surveillance. The combination of monomeric Ln-γ2 and prothrombin induced by Vitamin K Absence II (PIVKA-II) may be more sensitive for clinical diagnosis of HCC than any currently used combination

Expression of miR-21 is induced by signaling and high glucose conditions in RPECs.

<p>(A, B, and C) Results of quantitative (q) PCR analyses of miR-21, miR-204, and let-7e levels in TGF-β-stimulated ARPE-19 cells. Fold changes in expression levels of miR-21 (A), miR-204 (B), and let-7e (C) in TGF-β-stimulated ARPE-19 cells were determined relative to the unstimulated control cells. (D) Results of qPCR analyses of TGF-β-induced expression of miR-21 in ARPE-19 cells under low or high glucose conditions. Fold changes in expression levels of miR-21 in TGF-β-stimulated ARPE-19 cells under low or high glucose culture conditions were determined relative to the unstimulated control cells. RPECs, retinal pigment epithelial cells; TGF-β, transforming growth factor-β; LG = low glucose; HG = high glucose. *<i>*p</i> < 0.01; *<i>p</i> < 0.05.</p

Effect of miR-21 expression on cell proliferation of ARPE-19 cells.

<p>(A, B) Results of quantitative (q)PCR analyses of miR-21 in miR-21 mimic- or inhibitor-transfected ARPE-19 cells. Fold changes in expression levels of miR-21 in miR-21 mimic-transfected ARPE-19 cells were determined relative to the negative control mimic-transfected cells (A). Expression level of miR-21 in miR-21 inhibitor-transfected ARPE-19 cells in the presence or absence of TGF-β2 was calculated by comparing with negative control inhibitor-transfected cells (B). (C) Effect of miR-21 mimic or miR-21 inhibitor transfection on GFP reporter gene expression of CMV-EGFP or CMV-EGFP-3′ UTR miR-21-target reporter plasmid in ARPE19 cells. ARPE-19 cells were co-transfected with CMV-EGFP or CMV-EGFP-3′ UTR miR-21-target reporter plasmid, together with miR-21 mimics or miR-21 inhibitor, as indicated. After 24 h, GFP expression was analyzed under the fluorescence microscope. All fluorescence images were taken at the same exposure time (1/15s). (D, E) The migration ability of miR-21 mimic and miR-21 inhibitor-treated ARPE-19 cells was examined using the wound healing assay. (D) Representative images of wound sealing of miR-21 mimic and miR-21 inhibitor-transfected cells cultured with or without TGF-β at 0 and 12 h. (E) Quantification of the wound healing assay. The figure shows the average percentage of wound closure ± SD. The transfection of the miR-21 mimic significantly increased cell migration of ARPE-19 cells without TGF-β stimulation, and transfection of miR-21 inhibitor inhibited TGF-β-induced enhanced migration. (F, G) Cell viability was measured with CCK-8 kits following the treatment of miR-21 mimic and miR-21 inhibitor for 12, 24, 48, 72, and 96 h (F). The miR-21 mimic treatment resulted in increased cell proliferation, and miR-21 inhibitor treatment resulted in decreased cell proliferation at 72 h (G). NC, negative control. ***p < 0.001; **p < 0.01; *p < 0.05.</p

Effect of miR-21 expression on epithelial-mesenchymal transition (EMT)-related gene expression during TGF-β2-induced EMT of ARPE-19 cells.

<p>(A, B, and C) Results of qPCR analyses of fibronectin, αSMA, and N-cadherin mRNA expression in miR-21 mimic- or inhibitor-transfected ARPE-19 cells in the presence or absence of TGF-β2. Fold changes in expression levels of fibronectin (A), αSMA (B), and N-cadherin (C) mRNAs in miR-21 mimic-transfected ARPE-19 cells were determined relative to the untransfected control. NC, negative control; αSMA, alpha smooth muscle actin; EMT, epithelial-mesenchymal transition. ***p < 0.001; **p < 0.01; *p < 0.05; n.s; not significant.</p

Comparisons of expression levels of miR-21, miR-204, and let-7e in the vitreous humor of MH, RRD, PVR, and PDR samples.

<p>(A, B, and C) Results of quantitative PCR (qPCR) analyses of miR-21, miR-204, and let-7e levels from the vitreous humor of MH (n = 7), RRD (n = 5), PVR (n = 5), and PDR (n = 10) samples. Fold changes in expression levels of miR-21(A), miR-204 (B), and let-7e (C) in RRD, PVR, and PDR samples were determined relative to the control MH sample. CTR, control; RRD, rhegmatogenous retinal detachment; MH, macular hole; PVR, proliferative vitreoretinopathy; PDR, proliferative diabetic retinopathy. **<i>p</i> < 0.01; *<i>p</i> < 0.05.</p