LitCovid-sentences  

PMC:7019868 / 17447-23718 JSONTXT 11 Projects

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Id Subject Object Predicate Lexical cue
T127 0-2 Sentence denotes 3.
T128 3-10 Sentence denotes Results
T129 12-16 Sentence denotes 3.1.
T130 17-46 Sentence denotes Recovery of recombinant PEDVs
T131 47-284 Sentence denotes We have recently developed a reverse genetic platform of PEDV and have successfully generated a recombinant virus, iPEDVPT-P96, which is phenotypically comparable to its parental attenuated Taiwan PEDV-Pintung 52 strain, PEDVPT-P96 [19].
T132 285-587 Sentence denotes To study the role of the S gene in the attenuation mechanism of PEDVPT-P96, we further generated the recombinant, highly virulent Taiwan PEDVPT 52 strain, iPEDVPT-P5, and two chimeric viruses (iPEDVPT-P5-96S and iPEDVPT-P96-5S) by replacing the complete sequence of the S gene reciprocally (Figure S1).
T133 588-704 Sentence denotes In general, the approach to rescue the new recombinant viruses were virtually the same as published previously [19].
T134 705-940 Sentence denotes However, for the generation of iPEDVPT-P5 and iPEDVPT-P5-96S, we split the plasmid B to two fragments at nucleotide position of 9654 according to Hou et al. [23] in order to compensate for the instability caused by the toxic sequences.
T135 941-1103 Sentence denotes The in vitro and in vivo properties of iPEDVPT-P5 were confirmed similarly to those of the parental PEDVPT-P5 strain in a seven-day-old conventional piglet model.
T136 1104-1229 Sentence denotes At 24–48 h post-electroporation, typical PEDV cytopathic effects of giant syncytia were observed for all recombinant viruses.
T137 1230-1450 Sentence denotes Viral stocks were prepared by passaging viruses in Vero cells one additional time (P1), and the presence of PEDV was confirmed by detection of PEDV S protein by immunofluorescence assay (Figure 1A) and sequence analyses.
T138 1452-1456 Sentence denotes 3.2.
T139 1457-1503 Sentence denotes In Vitro Characterization of Recombinant PEDVs
T140 1504-1668 Sentence denotes The in vitro characteristics of the recombinant viruses were compared by evaluation of the size of syncytia, growth kinetics in Vero cells, and plaque morphologies.
T141 1669-1746 Sentence denotes All recombinant viruses induced formation of syncytia by 18 h post-infection.
T142 1747-1981 Sentence denotes Viruses carrying the identical S gene exhibited similar fusogenic ability as suggested by the comparable number of nuclei per syncytium; representative micrographs used to count the number of nuclei in syncytia are shown in Figure 1A.
T143 1982-2164 Sentence denotes Remarkably, viruses carrying S gene derived from iPEDVPT-P96, induced 3.5 to 4 times larger syncytia than the other two viruses containing S gene derived from iPEDVPT-P5 (Figure 1B).
T144 2165-2356 Sentence denotes Replication of all recombinant viruses in Vero cells were examined by performing both one-step and multistep growth kinetics at a multiplicity of infection (MOI) of 1 and 0.001, respectively.
T145 2357-2665 Sentence denotes Interestingly, although replacement of the S gene resulted in an alteration of the replication kinetics and efficiency, leading to similar growth curve patterns between viruses carrying the identical S gene sequence, it did not fully reverse the capability of virus yield, at least in Vero cells (Figure 1C).
T146 2666-2778 Sentence denotes We observed that plaque morphologies of the four recombinant viruses correlated to the results of syncytia size.
T147 2779-3095 Sentence denotes At 72 h post-infection, both iPEDVPT-P5 and iPEDVPT-P96-5S induced barely visible viral plaques in Vero cells macroscopically, whereas iPEDVPT-P96 and iPEDVPT-5-96S generated distinct and comparable viral plaques with the plaque size of iPEDVPT-P5-96S being slightly smaller than that of the iPEDVPT-P96 (Figure 1D).
T148 3096-3270 Sentence denotes These data suggested that the exchange of the S gene between iPEDVPT-P5 and iPEDVPT-P96 affected the fusogenicity and, to a lesser extent, replication kinetics in Vero cells.
T149 3272-3276 Sentence denotes 3.3.
T150 3277-3357 Sentence denotes Investigation of the Role of Spike Gene on the Pathogenicity of PEDVPT 52 strain
T151 3358-3644 Sentence denotes To evaluate the pathogenicity related to S gene replacement in PEDVPT 52 strain, we orally inoculated 2 mL of 5 × 102 TCID50/mL of iPEDVPT-P5, iPEDVPT-P5-96S, iPEDVPT-P96, iPEDVPT-P96-5S viruses and with PI medium in seven-day-old crossbred piglets assigned in the corresponding groups.
T152 3645-3771 Sentence denotes Parameters used to assess the pathogenicity of different recombinant viruses were summarized in Table 2 and shown in Figure 2.
T153 3772-3928 Sentence denotes The target sequences of the recombinant viruses at the time point of peak fecal viral shedding in each pig was confirmed identical to the original inoculum.
T154 3929-4051 Sentence denotes No viral RNA shedding, diarrhea or mortality was detected in the mock-treated group during the entire experimental course.
T155 4052-4357 Sentence denotes Piglets inoculated with recombinant viruses carrying the S gene derived from the highly virulent iPEDVPT-P5, namely the iPEDVPT-P5 itself and iPEDVPT-P96-5S, had an early onset of clinical symptoms including diarrhea, anorexia and decreased activity, and peak viral shedding at 1 d post-inoculation (DPI).
T156 4358-4624 Sentence denotes At 3 DPI, piglets inoculated with iPEDVPT-P5 (n = 3) and iPEDVPT-P96-5S (n = 3) both exhibited extensive PEDV-induced villous blunting and atrophy in the jejunum and, to a lesser extent, in the ileum; histological changes in the duodenum were not obvious (Figure 3).
T157 4625-4781 Sentence denotes Mortality rates of both groups were comparable, reaching 50% at 2 (iPEDVPT-P5 group) and 5 (iPEDVPT-P96-5S group) DPI and ultimately exceeded 80% by 10 DPI.
T158 4782-4985 Sentence denotes In comparison, inoculation with iPEDVPT-P5-96S and iPEDVPT-P96 viruses containing the S gene derived from the attenuated iPEDVPT-P96, induced a delayed onset of clinical symptoms and peak viral shedding.
T159 4986-5237 Sentence denotes Histological evaluation of piglets inoculated with both viruses (iPEDVPT-P96, n = 3; iPEDVPT-P5-96S, n = 3) at 3 DPI revealed a much milder degree of villous atrophy in the jejunum with conspicuous villous hyperplasia compared to the other two groups.
T160 5238-5393 Sentence denotes Notably, iPEDVPT-P5-96S-treated piglets exhibited statistically significant severer villous atrophy in jejunum than iPEDVPT-P96-treated piglets (Figure 3).
T161 5394-5695 Sentence denotes Despite the similar pattern and severity of viral shedding and clinical symptoms, inoculation with iPEDVPT-P5-96S eventually resulted in a higher mortality rate (40% in iPEDVPT-P96 and 80 % and iPEDVPT-P5-96S) and lower average daily weight gain by 10 DPI than those upon inoculation with iPEDVPT-P96.
T162 5696-5913 Sentence denotes These data suggested that iPEDVPT-P96 fully regained virulence by replacement of the S gene from the virulent PEDVPT 52 strains and the complementary approach can only partially reduce the virulence of the iPEDVPT-P5.
T163 5914-6185 Sentence denotes In agreement with the previous study, immunohistochemistry using anti-PEDV nucleocapsid monoclonal antibody demonstrated positive signals predominantly in the jejunal and ileal enterocytes at the top of villi (Figure 4) but occasionally within the mesenteric lymph nodes.
T164 6186-6271 Sentence denotes No difference in the viral distribution was identified among the recombinant viruses.