Patients’ characteristics and change in refractive errors of age-matched and spherical equivalent refractive error-matched groups of hyperopic patients who received full or undercorrection of hyperopia, with and without strabismus.

<p>Mean ± standard deviation (range); y = years; D = diopters; SER = spherical equivalent refractive error.</p>a<p>One-way ANOVA;</p>b<p>Linear-by-linear association.</p><p>Patients’ characteristics and change in refractive errors of age-matched and spherical equivalent refractive error-matched groups of hyperopic patients who received full or undercorrection of hyperopia, with and without strabismus.</p

Meta-Analysis of Cardiovascular Mortality with Use of Bisphosphonates.

<p>* Abbreviations: CI, confidence interval; CV, cardiovascular; HORIZON-PFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Pivotal Fracture Trial; HORIZON-RFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Recurrent Fracture Trial; M-H, Mantel Haenszel; OR, odds ratio. * Karam (2007) et al. included data from 6 trials of risedronate. Individual study data were not available.</p

Meta-Analysis of Myocardial Infarction and Stroke with Use of Bisphosphonates.

<p>* Abbreviations: CI, confidence interval; HORIZON-PFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Pivotal Fracture Trial; HORIZON-RFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Recurrent Fracture Trial; M-H, Mantel Haenszel; OR, odds ratio. * Karam (2007) et al. included data from 6 trials of risedronate. Individual study data were not available.</p

Meta-Analysis of Atrial Fibrillation Associated with Use of Bisphosphonates.

<p>* Abbreviations: CI, confidence interval; EPIC, Early Postmenopausal Intervention Cohort study; FIT, Fracture Intervention Trial; FOSIT, Fosamax International Trial; HORIZON-PFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Pivotal Fracture Trial; HORIZON-RFT, the Health Outcomes and Reduced Incidence with Zoledronic Acid Once Yearly Recurrent Fracture Trial; M-H, Mantel Haenszel; OR, odds ratio. * Lewiecki (2010) et al. included data from 4 trials of ibandronate. Karam (2007) et al. included data from 6 trials of risedronate. Individual study data were not available.</p

Selection of Randomized Controlled Trials of Bisphosphonates for Meta-Analysis.

<p>Abbreviations: BIS, bisphosphonate; CTR, control; HIV, human immunodeficiency virus; RCT, randomized controlled trial</p

Meta-Analysis of Total Adverse Cardiovascular Events Associated with Use of Bisphosphonates.

<p>Abbreviations: CI, confidence interval; CV, cardiovascular; EPIC, Early Postmenopausal Intervention Cohort study; IVF, IntraVenous Fracture study; M-H, Mantel Haenszel; OR, odds ratio; VERT-MN, Vertebral Efficacy with Risedronate Therapy Multinational Study</p

A Nucleic-Acid Hydrolyzing Single Chain Antibody Confers Resistance to DNA Virus Infection in HeLa Cells and C57BL/6 Mice

<div><p>Viral protein neutralizing antibodies have been developed but they are limited only to the targeted virus and are often susceptible to antigenic drift. Here, we present an alternative strategy for creating virus-resistant cells and animals by ectopic expression of a nucleic acid hydrolyzing catalytic 3D8 single chain variable fragment (scFv), which has both DNase and RNase activities. HeLa cells (SCH7072) expressing 3D8 scFv acquired significant resistance to DNA viruses. Virus challenging with Herpes simplex virus (HSV) in 3D8 scFv transgenic cells and fluorescence resonance energy transfer (FRET) assay based on direct DNA cleavage analysis revealed that the induced resistance in HeLa cells was acquired by the nucleic acid hydrolyzing catalytic activity of 3D8 scFv. In addition, pseudorabies virus (PRV) infection in WT C57BL/6 mice was lethal, whereas transgenic mice (STG90) that expressed high levels of 3D8 scFv mRNA in liver, muscle, and brain showed a 56% survival rate 5 days after PRV intramuscular infection. The antiviral effects against DNA viruses conferred by 3D8 scFv expression in HeLa cells as well as an <i>in vivo</i> mouse system can be attributed to the nuclease activity that inhibits viral genome DNA replication in the nucleus and/or viral mRNA translation in the cytoplasm. Our results demonstrate that the nucleic-acid hydrolyzing activity of 3D8 scFv confers viral resistance to DNA viruses <i>in vitro</i> in HeLa cells and in an <i>in vivo</i> mouse system.</p></div

3D8 scFv inhibits HSV-1 encoded gene expression by DNase and RNase activity.

<p><b>A</b>. Schematic diagram for identification of viral gene expression pattern by HSV-1 challenging. <b>B</b>. Wild-type HeLa, SCH07072, and muSCH cells were infected with HSV::GFV at an MOI of 0.5 and then incubated for 2, 6.5, and 25 hr in the presence or absence of PAA 1 hr after virus challenge. ICP4 was used for the immediate early stage, UL9 for the early stage, and UL19 for the late stage of viral infection. Data bars show mean ± standard error. ** and *** indicate significant differences from HSV-1 viral DNA at <i>p<0.01</i> and <i>p<0.001</i>, respectively (one-way analysis of variance and Tukey's post hoc <i>t</i>-test). <b>C</b>. Quantitative real-time reverse transcription-polymerase chain reaction (RT-PCR) was used to measure the expression of immediate early genes (ICP0 and ICP4), early genes (UL9 and UL29), and late genes (UL19 and UL38). The relative concentrations of HSV mRNAs were calculated after normalization to the GAPDH gene using the delta delta C_T method. Data bars represent mean ± standard error. *, **, *** indicate significant differences from HeLa cells at <i>p<0.05, p<0.01</i>, and <i>p<0.001</i>, respectively (one-way analysis of variance and Tukey's post hoc <i>t</i>-test).</p

STG90 exhibits antiviral effects against PRV.

<p><b>A</b>. The expression levels of PRV gpD RNA in the muscle and brain of WT-Mock, WT-PRV, STG69-PRV, STG90-PRV, and STG135-PRV mice were investigated. Only live STG90-PRV mice did not show PRV gpD expression. <b>B</b>. Immunohistochemistry was performed to detect PRV in PRV target organs; brain (upper panel: ×100) and femoral muscle (bottom panel: ×400). The PRV gpD protein was stained with a monoclonal anti-PRV antibody and visualized with DAB. The Purkinje layer cells in the WT-PRV group stained brown for the gpD protein, whereas no staining was observed in the other groups.</p

3D8 scFv cannot hydrolyze methylated DNA or protein-bound DNA.

<p><b>A</b>. HeLa methylated genomic DNA and non-methylated DNA (NEB) were treated with 3D8 scFv and DNase I at four concentrations: 10×10⁻⁴ U/µl, 8.3×10⁻⁴ U/µl, 1.4×10⁻⁴ U/µl, and 0 U/µl. The DNA samples were harvested and analyzed by electrophoresis at 0, 1, 2, and 3 hr after treatment. At a concentration of 8.3×10⁻⁴ U/µl, 3D8 scFv digested more non-methylated DNA than DNase I 1 hr after treatment. <b>B</b>. Chromatin was prepared using the EZ-Zyme Enzymatic Chromatin Prep kit. Prepared chromatin DNA and naked DNA were treated with 3D8 scFv and DNase I (10×10⁻⁴ U, 8.3×10⁻⁴ U, and 0 U/µl). The DNA samples were harvested and analyzed by electrophoresis at 0, 1, 2, and 3 hr after treatment. Neither 3D8 scFv nor DNase I hydrolyzed histone-bound DNA at concentrations of 10×10⁻⁴ U/µl and 8.3×10⁻⁴ U/µl. When naked DNA was treated with 3D8 scFv at concentrations of 8.3×10⁻⁴ U/µl, 3D8 scFv could digest naked DNA. But, DNase I could not digest naked DNA at 8.3×10⁻⁴ U/µl. <b>C</b>. The relative quantitative values of each DNA of methylated, non-methylated, histone-bound, and naked HeLa genomic DNA were measured for 3 hr after each DNA was treated with 3D8 scFv (8.3×10⁻⁴ U/µl). The bar graphs indicate the average values of each sample calculated from three individual experiments.</p