Detection of Cartilage Oligomeric Matrix Protein Using a Quartz Crystal Microbalance

Current methods for diagnosing early stage osteoarthritis (OA) based on the magnetic resonance imaging and enzyme-linked immunosorbent assay methods are specific, but require specialized laboratory facilities and highly trained personal to obtain a definitive result. In this work, a user friendly and non-invasive quartz crystal microbalance (QCM) immunosensor method has been developed to detect Cartilage Oligomeric Matrix Protein (COMP) for early stage OA diagnosis. This QCM immunosensor was fabricated to immobilize COMP antibodies utilizing the self-assembled monolayer technique. The surface properties of the immunosensor were characterized by its FTIR and electrochemical impedance spectra (EIS). The feasibility study was based on urine samples obtained from 41 volunteers. Experiments were carried out in a flow system and the reproducibility of the electrodes was evaluated by the impedance measured by EIS. Its potential dynamically monitored the immunoreaction processes and could increase the efficiency and sensitivity of COMP detection in laboratory-cultured preparations and clinical samples. The frequency responses of the QCM immunosensor changed from 6 kHz when testing 50 ng/mL COMP concentration. The linear regression equation of frequency shift and COMP concentration was determined as: y = 0.0872 x + 1.2138 (R2 = 0.9957). The COMP in urine was also determined by both QCM and EIS for comparison. A highly sensitive, user friendly and cost effective analytical method for the early stage OA diagnosis has thus been successfully developed

[[alternative]]以律令為中心

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氣候、災荒與生存抉擇--唐代鬻賣人口現象析論—唐代鬻賣人口現象析論

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The relationship between optic nerve head deformation and visual field defects in myopic eyes with primary open-angle glaucoma.

PurposeTo investigate the relationship between the morphologic features of myopic optic nerve head (ONH) and visual field (VF) defects in myopic subjects with primary open-angle glaucoma (POAG) by intraindividual comparison.MethodsMyopic POAG subjects with unilateral glaucomatous VF defect were recruited. The morphologic features of myopic ONH, including optic disc tilt, optic disc rotation, and β-zone parapapillary atrophy (PPA) were measured from color fundus photographs. The comparisons were performed between the eyes with VF defects and the contralateral eyes without VF defects. Logistic regression analysis was performed to investigate the relationship between various ocular parameters and the presence of VF defects.ResultsWe retrospectively included 100 eyes of 50 myopic POAG subjects. (Mean age: 50.1 ± 10.0 years). The tilt ratio was similar between the paired eyes. The degree of optic disc rotation (12.96 ± 7.21°) in eyes with VF defects were statistically greater than the contralateral eyes (6.86 ± 4.30°; P ConclusionsAmong the morphologic features of myopic ONH, only the greater degree of the optic disc rotation was associated with the presence of VF defects in myopic subjects with POAG

BRD4 Phosphorylation Regulates HPV E2-Mediated Viral Transcription, Origin Replication, and Cellular MMP-9 Expression

Post-translational modification can modulate protein conformation and alter binding partner recruitment within gene regulatory regions. Here, we report that bromodomain-containing protein 4 (BRD4), a transcription co-factor and chromatin regulator, uses a phosphorylation-induced switch mechanism to recruit E2 protein encoded by cancer-associated human papillomavirus (HPV) to viral early gene and cellular matrix metalloproteinase-9 (MMP-9) promoters. Enhanced MMP-9 expression, induced upon keratinocyte differentiation, occurs via BRD4-dependent recruitment of active AP-1 and NF-κB to their target sequences. This is triggered by replacement of AP-1 family members JunB and JunD by c-Jun and by re-localization of NF-κB from the cytoplasm to the nucleus. In addition, BRD4 phosphorylation is critical for E2- and origin-dependent HPV DNA replication. A class of phospho-BRD4-targeting compounds, distinct from the BET bromodomain inhibitors, effectively blocks BRD4 phosphorylation-specific functions in transcription and factor recruitment

The Influence of the Degradation of Tetracycline by Free Radicals from Riboflavin-5′-Phosphate Photolysis on Microbial Viability

Tetracycline (TC) is a broad-spectrum antibiotic compound. Wastewater with TC may have an adverse effect on ecosystems. Riboflavin-5′-phosphate (FMN or flavin mononucleotide) is a non-toxic product of the phosphorylation of vitamin B2 and is required for the proper functioning of the humans. FMN is sensitized to ultraviolet (UV) and blue light radiation, as evidenced by the generation of reactive oxygen species (ROS). This study inspects feasible applications of blue light on FMN so as to develop a valid way of degrading TC by FMN photolysis. We used the increased rate of bacterial survival as a practical indicator of antibiotic degradation. TC in the presence of FMN solution decomposed completely after 20 W/m2 of blue light irradiation (TCF treatment), and the degradation of TC (D-TCF) occurred after the photolytic process. After TCF treatment, colony-forming units (CFUs) of Escherichia coli (E. coli) were determined for the D-TCF solution. The CFU of E. coli preservation was 93.2% of the D-TCF solution (50 μg/mL of TC in the presence of 114 μg/mL of FMN solution treated with 20 W/m2 of blue light irradiation at 25 °C for 1 h) cultivation. The mass spectrum of D-TCF showed diagnostic ion signals at m/z 431.0 and 414.0 Da. The molecular formula of D-TCF was C21H22N2O8, and the exact mass was 430.44 g/mol. TC degradation by FMN photolysis can significantly decrease the antimicrobial ability of TC. The results expressed here regarding the influence of FMN photolysis on TC degradation offer an environmentally sound wastewater treatment method

Brd4 links chromatin targeting to HPV transcriptional silencing

The E2 protein encoded by human papillomaviruses (HPVs) inhibits expression of the viral E6 oncoprotein, which, in turn, regulates p53 target gene transcription. To identify cellular proteins involved in E2-mediated transcriptional repression, we isolated an E2 complex from human cells conditionally expressing HPV-11 E2. Surprisingly, the double bromodomain-containing protein Brd4, which is implicated in cell cycle control and viral genome segregation, was found associated with E2 and conferred on E2 the ability to inhibit AP-1-dependent HPV chromatin transcription in an E2-binding site-specific manner as illustrated by in vitro reconstituted chromatin transcription experiments. Knockdown of Brd4 in human cells alleviates E2-mediated repression of HPV transcription. The E2-interacting domain at the extreme C terminus and the chromatin targeting activity of a bromodomain-containing region are both essential for the corepressor activity of Brd4. Interestingly, E2–Brd4 blocks the recruitment of TFIID and RNA polymerase II to the HPV E6 promoter region without inhibiting acetylation of nucleosomal histones H3 and H4, indicating an acetylation-dependent role of Brd4 in the recruitment of E2 for transcriptional silencing of HPV gene activity. Our finding that Brd4 is a component of the virus-assembled transcriptional silencing complex uncovers a novel function of Brd4 as a cellular cofactor modulating viral gene expression

qPCR primer sequences.

<p>qPCR primer sequences.</p

CoCl₂ induces abnormal neovascularization in zebrafish embryos.

<p>(A) Morphological images obtained by an optical microscope revealed no severe phenotype in CoCl_<b>2</b>-treated Tg(<i>fli1a</i>:<i>EGFP</i>) embryos at 3 dpf. (B) With fluorescence excitation, ectopic SIV and excessive retinal vascularization are shown in CoCl_<b>2</b>-treated embryos compared with the untreated control. (C, D) A dose-dependent decrease in survival rates (four independent experiments; n = 35 in each group) in embryos treated with increasing concentrations (0–20 mM) and an increase in the vascular defect occurrence rate in the SIV and retinal vessels are shown (three independent experiments; n = 35 in each group). (E, F) Real-time RT-PCR data show that CoCl_<b>2</b> treatment causes overexpression of <i>vegfaa</i> and <i>vegfr2</i> mRNAs in zebrafish. (G,H) Absolute quantification of the copy number of <i>vegfr2</i> and <i>plxnd1</i> by real-time PCR are reduced at 5 dpf versus 3 dpf. Each bar represents the mean ± SEM. * (<i>p</i> < 0.01) and ** (<i>p</i> < 0.001) compared with the mock control group. SIV, subintestinal vessel; RV, retinal vessel.</p