Yes / Laboratory experiments are presented to investigate the effect of different piggyback pipeline configurations on the morphology of local scour under wave conditions. Scour depth and width around the pipelines under regular and irregular waves are measured and analyzed for a range of pipeline and wave conditions; such as the spacing between two pipes (G), gap between the main pipe and seabed (e), pipe diameter (D), wave height (H) and period (T). Experimental results reveal that both the scour depth and width around piggyback pipeline is much larger than those around single pipe under the same wave conditions. Scour depth increases with the increase of the Keulegan-Carpenter (KC) number and decreases with increase of G and e. When e exceeds 0.5D, scour depth tends to approach 0.When spacing G is greater than 0.4D, the destabilization from small pipe to large one is greatly reduced, resulting in scour depth around piggyback pipeline being close to that around single pipe. Similar to scour depth, scour width broadens with the increase of KC number increasing and decreases with the increase of G. Experiments also show that the effect of e on scour depth is greater than that of G under the same test conditions, while their impact on scour width is opposite. Furthermore, scour width under irregular waves is extended slightly compared with regular wave for otherwise the identical conditions. / National Natural Science Foundation (No. 51279189).

Yes / This paper presents the results from a numerical simulation study to investigate the effect of the position angle (α) of small pipeline on the local scour and the hydrodynamic force around the piggyback pipeline in steady current conditions. Results show that the local scour depth around the piggyback pipeline increases first and then decreases with the increase of α. The scour depth and width reach the maximum values as the small pipe locates at the top of the large pipeline (i.e. α = 90°). The scour around the piggyback pipeline is accelerated when α ranges between 30° and 165°, while for α = 0°–30° and 165°–180°, the local scour around the piggyback pipeline is inhibited. Furthermore, the small pipe placed in front of the large pipe has slightly larger effect on the scour hole morphology than that when it is placed behind the large pipe. The drag force coefficient increases first and reaches the maximum value at α = 75°, and then decreases with the increase of α. Eventually the drag force coefficient approaches roughly a constant. The lift force coefficient is approximately a V-shaped with the variation of α and has the maximum value at α = 90°.

Secondary infusions is a common method to deliver short infusions of intravenous (IV) drugs and fluids. Errors associated with this infusion method have led to patient safety concerns. This study's objective was to empirically evaluate interventions to mitigate secondary infusion risks. Three interventions, including a technology-based intervention (clamp detector on a smart pump), a training-based intervention (educational module), and a practice-based intervention (use of a separate pump for short infusions), were tested in a simulated inpatient unit. The technology-based intervention significantly decreased secondary clamp errors whereas the training-based intervention reduced complex pressure differential errors. The practice-based intervention was the only intervention that significantly decreased both secondary clamp errors and pressure differential errors, but introduced new risks due to mismanagement of residual volume in IV tubing. Study results highlight the need for a combination of mitigation strategies and can help guide the selection of interventions to reduce secondary infusion errors.

Secondary infusions is a common method to deliver short infusions of intravenous (IV) drugs and fluids. Errors associated with this infusion method have led to patient safety concerns. This study's objective was to empirically evaluate interventions to mitigate secondary infusion risks. Three interventions, including a technology-based intervention (clamp detector on a smart pump), a training-based intervention (educational module), and a practice-based intervention (use of a separate pump for short infusions), were tested in a simulated inpatient unit. The technology-based intervention significantly decreased secondary clamp errors whereas the training-based intervention reduced complex pressure differential errors. The practice-based intervention was the only intervention that significantly decreased both secondary clamp errors and pressure differential errors, but introduced new risks due to mismanagement of residual volume in IV tubing. Study results highlight the need for a combination of mitigation strategies and can help guide the selection of interventions to reduce secondary infusion errors.

Chez les plantes, le RNA silencing (RNAi) est le principal mécanisme de défense antivirale. Il est opéré par de petites molécules d’ARN (siRNA), de 21-22nt de long, générées à partir de l’ARN viral par DCL4 et DCL2, respectivement. Ces siRNA confèrent la séquence-spécificité des réactions de défense intracellulaire et peuvent se déplacer à longue distance pour immuniser les cellules saines. En conséquence, les virus ont développé des protéines (VSRs) capables de supprimer ces deux aspects du RNAi. Au cours de cette thèse, j’ai pu démontrer in vivo que la protéine P15 du Peanut clump virus (PCV) est capable de séquestrer les siRNA de 21 et 22nt et qu’elle bloque le mouvement de ces derniers plus efficacement que ceux de 21nt. Pour compenser cette faiblesse, au cours de l’infection par le PCV, P15 est transportée à l’intérieur des peroxisomes en association avec les siRNA qu’elle séquestre. Le confinement des siRNA mobiles de 21nt à l’intérieur de ces organelles conduit à une inhibition du RNAi systémique et stimule fortement la propagation du PCV à travers la plante. Ces travaux définissent une nouvelle stratégie de pathogénèse virale au cours de laquelle une organelle est utilisé pour neutraliser des molécules de défense produites par l’hôte. / In plants, RNA interference (RNAi) is the main antiviral defense mechanism. It is initiated through the processing of viral RNA into 21-22nt long siRNA by DCL4 and DCL2, respectively. These siRNA can mediate sequence-specific local defense reactions (cell-autonomous RNAi) or move to distant tissues to prime defenses in naive cells (systemic RNAi). Consequently, viruses have evolved proteins (VSRs) to suppress both aspects of RNAi. In this in vivo study, I show that P15, the VSR of Peanut clump virus (PCV), binds and sequesters both 21nt and 22nt siRNA. Importantly, it stops the movement of 22nt siRNA more efficiently than 21nt siRNA. During infection, P15 is shuttled into peroxisomes, and is able to « piggyback » siRNA into these organelles. By confining mobile DCL4-dependent antiviral 21nt siRNA within peroxisomes, P15 is able to shut down systemic RNAi and strongly promote PCV movement. This work describes a novel pathogenic strategy in which an organelle is used to neutralize host defensive molecules.

Factor VIII is a clotting factor found on the intrinsic side of the coagulation cascade. A mutation in the factor VIII gene causes the disease Hemophilia A, for which there is no cure. The most common treatment is administration of recombinant factor VIII. However, this can cause an immune response that renders the treatment ineffective in certain hemophilia patients. For this reason a new treatment, or cure, needs to be developed. Gene editing is one solution to correcting the factor VIII mutation. CRISPR/Cas9 mediated gene editing introduces a double stranded break in the genomic DNA. Where this break occurs repair mechanisms cause insertions and deletions, or if a template oligonucleotide can be provided point mutations could be introduced or corrected. However, to accomplish this goal for editing factor VIII mutations, a way to deliver the components of CRISPR/Cas9 into somatic cells is needed. In this study, I confirmed that the CRISPR/Cas9 system was able to create a mutation in the factor VIII gene in zebrafish. I also showed that the components of CRISPR/Cas9 could be piggybacked by vivo morpholino into a variety of blood cells. This study also confirmed that the vivo morpholino did not interfere with the gRNA binding to the DNA, or Cas9 protein inducing the double stranded break.

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In the past couple of decades, the zebrafish has been widely used to study hemostatic disorders. In this study, we generated a CRISPR/Cas9 mediated zebrafish mutant that contains a 55-nucleotide insertion in exon 29 of the von Willebrand factor (vwf) gene. The mutants had impaired ristocetin-mediated agglutination of whole blood, prolonged PTT and more bleeding in the lateral incision compared to wild-type fish. The bleeding phenotype observed here is similar to the phenotype observed in vwf knockout mice and patients with von Willebrand disease (VWD). The mutant model developed here can thus be used for exploring the role of Vwf in angiogenesis and for developing gene therapy. The deficiency of VWF causes VWD and the etiology remains unknown in 30% of Type 1 VWD cases. Previous studies have identified that the ABO blood group and ST3GAL4 (glycosyltransferases) are involved in the regulation of VWF levels. Since VWF is heavily glycosylated, we hypothesized that other glycosyltransferases may also be involved in regulating VWF. We performed a knockdown screen of 234 glycosyltransferase genes and identified 14 genes that altered Vwf levels. The sequencing of these genes in Type 1 VWD patients could help identify novel mutations to decipher the molecular basis for the unknown etiologies in Type 1 VWD. Moreover, therapeutic interventions could be designed in the future by modulation of these gene products to control bleeding or thrombosis.Zebrafish has three f9 genes, f9a, f9b, and f9l and the ortholog to human F9 is unknown. RNA analysis showed an age-dependent increase in expression of all three genes from larval stages to adults, comparable to those observed in mice and humans while mass spectrometry and immunohistochemistry confirmed the presence of all three proteins in the fish. Based on coagulation assays performed after individual gene knockdown and immunodepletion, we identified that zebrafish f9a has functional activity similar to human F9 and Fixl is functionally similar to Fx. Thus, the zebrafish could be used to identify factors controlling f9 gene expression with age and for modeling Hemophilia B in the quest to develop gene therapy protocols. In zebrafish, dilute plasma with exogenously added human fibrinogen was used for kinetic coagulation assays. Here, we developed a microkinetic assay using 25% zebrafish or 30% human plasma followed by the addition of coagulation activators and CaCl2. Our results showed both zebrafish and human plasmas yielded kinetic PT, kinetic PTT, and kinetic Russel's viper venom time curves similar to previously established human kinetic curves. Moreover, clotting times derived from these kinetic curves were identical to human PT, PTT, and Russel's viper venom time. Thus, the microkinetic assay developed here could measure blood coagulation activity in small animal models like zebrafish and human blood samples obtained from a finger prick in adults or heel prick in infants.