Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Type A asparagine synthetase in the zoonotic Cryptosporidium parvum (CpAsnA): Biochemical features and potential as a novel therapeutic target

doi: 10.1016/j.ijpddr.2025.100601

Figure Lengend Snippet: Reactions and types of asparagine synthetase (ASNS) in mammals, Cryptosporidium parvum and other apicomplexans. (A) Comparison of the asparagine synthesis between mammalian hosts and C. parvum . Mammals can synthesize aspartate de novo from the glycolytic intermediate oxaloacetate by glutamic-oxaloacetic transaminase (GOT) and convert aspartate to asparagine by type B ASNS (ASNS-B or AsnB) that uses glutamine as the nitrogen donor. Asparagine can be broke down to aspartate and ammonia by asparaginase (ASPG). However, C. parvum cannot synthesize aspartate de novo, but possesses a standalone type A ASNS (CpAsnA) synthesizes asparagine from aspartate and ammonia. (B) The presence or absence of ASNS and types of ASNS in the major apicomplexan lineages based datamining the NCBI protein databases. The dinoflagellates and ciliates are included for comparison.

Article Snippet: Multi-enzyme coupled colorimetric assay was performed in 50 μL Tris-HCl buffer (100 mM; pH 7.5) containing 100 mM KCl and 10 mM MgCl 2 , mixed with the following substrates or cofactor: aspartate (3 mM), NH 4 Cl (10 mM), ATP (2 mM) and phosphoenolpyruvate (PEP; 2 mM) and NADH (1 mM), and the following recombinant enzymes: C. parvum pyruvate kinase (CpPK) (10 μg), C. parvum lactate dehydrogenase (CpLDH) (4 μg), and adenylate kinase (ADK; 4 units) (MilliporeSigma, Burlington, MA, USA).

Techniques: Comparison

Journal: International Journal for Parasitology: Drugs and Drug Resistance

Article Title: Type A asparagine synthetase in the zoonotic Cryptosporidium parvum (CpAsnA): Biochemical features and potential as a novel therapeutic target

doi: 10.1016/j.ijpddr.2025.100601

Figure Lengend Snippet: In vitro anti-cryptosporidial efficacy and cytotoxicity of the four top hits (CpAsnA inhibitors). (A) Illustration of the chemical structures and CAS numbers of the four CpAsnA inhibitors. (B) Cytotoxicity of the four inhibitors on the host cells (HCT-8 cells) as determined by determining the cellular ATP contents with a luminescent assay. The 50 % toxic concentrations ( TC 50 values) were much higher than 100 μM, while that of HATi-II was 91 μM. (C) In vitro efficacy of the specified three CpAsnA inhibitors against the growth of C. parvum in vitro as determined by a 44 h infection assay. The parasite loads were determined by qRT-PCR as described in the Materials and Methods. Assays were performed at least three times, and the data shown here were from a representative experiment. The error bars represent standard errors of the means (SEMs).

Article Snippet: Multi-enzyme coupled colorimetric assay was performed in 50 μL Tris-HCl buffer (100 mM; pH 7.5) containing 100 mM KCl and 10 mM MgCl 2 , mixed with the following substrates or cofactor: aspartate (3 mM), NH 4 Cl (10 mM), ATP (2 mM) and phosphoenolpyruvate (PEP; 2 mM) and NADH (1 mM), and the following recombinant enzymes: C. parvum pyruvate kinase (CpPK) (10 μg), C. parvum lactate dehydrogenase (CpLDH) (4 μg), and adenylate kinase (ADK; 4 units) (MilliporeSigma, Burlington, MA, USA).

Techniques: In Vitro, Luminescence Assay, Infection, Quantitative RT-PCR

Journal: Life Science Alliance

Article Title: STAT1-IFITM3 promotes autophagy in epithelial cells to control Cryptosporidium parvum infection

doi: 10.26508/lsa.202503200

Figure Lengend Snippet: Standard curves for C. parvum oocysts.

Article Snippet: C. parvum oocysts were purchased from Waterborne Inc. An appropriate amount of C. parvum oocysts was immersed in ice-cold PBS solution containing 20% sodium hypochlorite, pretreated for 10 min at 4°C, and centrifuged at 3, 667 g for 10 min.

Techniques: