Analysis of the subcellular localization of the proteins Rep, Rep' and Cap of porcine circovirus type 1

Porcine circovirus type 1 (PCV1) encodes two major ORFs. The cap gene comprises the major structural protein of PCV, the rep gene specifies Rep and Rep', which are both essential for initiating the replication of the viral DNA. Rep corresponds to the full-length protein, whereas Rep' is a truncated splice product that is frame-shifted in its C-terminal sequence. In this study, the cellular localization of PCV1-encoded proteins was investigated by immune fluorescence techniques using antibodies against Rep, Rep' and Cap and by expression of viral proteins fused to green and red fluorescence proteins. Rep and Rep' protein co-localized in the nucleus of infected cells as well as in cells transfected with plasmids expressing Rep and Rep' fused to fluorescence proteins, but no signal was seen in the nucleoli. Rep and Rep' carry three potential nuclear localization signals in their identical N-termini, and the contribution of these motifs to nuclear import was experimentally dissected. In contrast to the rep gene products, the localization of the Cap protein varied. While the Cap protein was restricted to the nucleoli in plasmid-transfected cells and was also localized in the nucleoli at an early stage of PCV1 infection, it was seen in the nucleoplasm and the cytoplasm later in infection, suggesting that a shuttling between distinct cellular compartments occurs.     

Ultrasonography and erythrocyte distribution width in patients with plantar fasciitis

BackgroundThe measurement of plantar fascia thickness with ultrasonography can be used for both for diagnosis and as a response-to-treatment parameter in plantar fasciitis. Furthermore, with the recent studies, red cell distribution width may be used as an inflammatory marker. Aim of this study is to investigate the association of red cell distribution width and ultrasonography on diagnosis and monitoring of treatment in patients with plantar fasciitis.MethodsClinically diagnosed 102 patients with plantar fasciitis between the dates January 2016 to July 2018 were analysed. Hemogram, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and plantar fascial ultrasonography were obtained on initial evaluation and in 1 month, 2 months and 3 months of the standard nonoperative treatment; American Orthopaedic Foot & Ankle Hindfoot Score (AOFAS) and Visual Analog Scale (VAS) scores were recorded. Posthoc and multivariate logistic regression analysis were used for statistical analysis on SPSS 21.0.ResultsRed cell distribution width was correlated with plantar fascia thickness by the end of the 1 month (r = 0.26, P = .013). Female sex, BMI over 30 kg/m², higher red cell distribution width and higher plantar fascia thickness were associated with plantar fasciitis on initial evaluation. Higher red cell distribution width together with higher plantar fascia thickness were also found to be a risk factor for both on initial evaluation and 1 month after treatment in plantar fasciitis.ConclusionThis study shows that association of red cell distribution width and plantar fascia thickness can be not only a diagnostic predictor but also an indicator of treatment response in plantar fasciitis.Level of clinical evidenceLevel IV     

Development of a Novel Human Cell-Derived Tissue-Engineered Heart Valve for Transcatheter Aortic Valve Replacement: an In Vitro and In Vivo Feasibility Study

Transcatheter aortic valve replacement (TAVR) is being extended to younger patients. However, TAVR-compatible bioprostheses are based on xenogeneic materials with limited durability. Off-the-shelf tissue-engineered heart valves (TEHVs) with remodeling capacity may overcome the shortcomings of current TAVR devices. Here, we develop for the first time a TEHV for TAVR, based on human cell-derived extracellular matrix and integrated into a state-of-the-art stent for TAVR. The TEHVs, characterized by a dense acellular collagenous matrix, demonstrated in vitro functionality under aortic pressure conditions (n?=?4). Next, transapical TAVR feasibility and in vivo TEHV functionality were assessed in acute studies (n?=?5) in sheep. The valves successfully coped with the aortic environment, showing normal leaflet motion, free coronary flow, and absence of stenosis or paravalvular leak. At explantation, TEHVs presented full structural integrity and initial cell infiltration. Its long-term performance proven, such TEHV could fulfill the need for next-generation lifelong TAVR prostheses.