CHA2DS2-VASc-HSF scores as a predictor of short-term clinical outcomes in ST-segment elevation myocardial infarction patients undergoing primary percutaneous coronary intervention

Figures

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  Fig. 1. Illustration of pyrimidine biosynthesis and purine salvage pathways in T. gondii. CPSII, carbamoyl phosphate synthetase II; ATC, aspartate transcarbamoylase; DHO, dihydroorotase; DHODH, dihydroorotate dehydrogenase; OPRT, orotate phosphoribosyltransferase; OMPDC, orotidine-5’-monophosphate decarboxylase; UMP, uridine-5’-monophosphate; UPRT, uracil phosphoribosyltransferase; UP, uridine phosphorylase; CTD, cytidine deaminase; HXGPRT, hypoxanthine-xanthine-guanine phosphoribosyltransferase; AK, adenosine kinase

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  Fig. 2. hENT1 inhibitor NBMPR impairs growth of pyrimidine auxotrophic parasites by blocking uracil transport. (A) Intracellular replication assay comparing parasite proliferation under the indicated condition (± uracil). Following synchronized invasion of HFF cells, parasites were allowed to proliferate for 24 h. The number of parasites (1, 2, 4, 8, ≥ 16) per parasitophorous vacuole was then quantified by IFA. Data are represented as the means ± SEM from three independent experiments. The statistical significance of differences in Tg/PV numbers across groups was assessed using two-way ANOVA, *0.01 ≤ p < 0.05, **0.001 ≤ p < 0.01, ***p < 0.001, NS: not significant. (B) Plaque assay comparing the growth of RHΔdho and RHΔatc parasites with or without 250 µM uracil supplementation to that of parental strain RH. Two hundred purified tachyzoites were used to infect HFFs for 7 days and then stained by 0.1% crystal violet. (C) Relative plaque sizes (calculated by Photoshop) formed by RHΔdho and RHΔatc strains versus RH strain from panel B. Data represent means ± SEM. Statistical significance was determined by one-way ANOVA, ***p < 0.001. (D) Model of nucleoside/nucleobase transport dynamics upon…

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  Fig. 3. Phylogenetic analyses of nucleoside/nucleobase transporter candidates in T. gondii. (A) Phylogenetic analysis of T. gondii nucleoside/nucleobase transporters with hENTs and hCNTs. Evolutionary relationships have been done in MEGA 7 using the neighbor-joining algorithm. (B) Multiple sequence alignment of TgENT1 and hENTs. The transmembrane domains (TMs), glycosylated loop and cytoplasmic loop are indicated based on the known features of hENT1 and hENT2 (Yao et al. 2002). (C) Phylogenetic placement of T. gondii nucleoside/nucleobase transporters within a broader context of uracil transporters from diverse taxa. (D) Multiple sequence alignment of TgENT1 and uracil transporters. All sequences of nucleoside and nucleobase transporters are retrieved from Uniprot (https://www.uniprot.org) and VEuPathDB (https://veupathdb.org/)

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  Fig. 4. Structure prediction and molecular docking analysis of TgENT1. (A) Whole-structure superposition analysis of hENT1 and TgENT1. The structure of hENT1 (PDB ID: 6OB6; Uniprot ID: Q99808; green) and TgENT1 (AlphaFold-predicted structure; Uniprot ID: S8F309; light blue) is performed using the PyMOL3.1. (B) Comparative structural analysis of hENT1 and TgENT1, focusing on the NBMPR-binding residues within 6Å. (C) The predicted binding pose of NBMPR in TgENT1, obtained using the Schrödinger suite, with a 2D interaction diagram showing key contacts. (D) Predicted binding poses of TgENT1 with nucleoside/nucleobase substrates: uridine, adenosine, adenine, and uracil

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  Fig. 5. Localization analysis of TgENT1. (A) Schematic illustration of endogenous C-terminal tagging of TgENT1 with a mAID-HA tag via CRISPR/Cas9-mediated homologous recombination. (B) Immunofluorescent staining using mouse-derived anti-SAG1 and rabbit-derived anti-HA antibodies to assess the expression and subcellular localization of TgENT1 in TgENT1-mAID parasites. Scale bar, 5 μm