The apicoplast is important for the viability and persistence of Toxoplasma gondii bradyzoites.

Toxoplasma gondii is responsible for toxoplasmosis, a disease that can be serious when contracted during pregnancy, but can also be a threat for immunocompromised individuals. Acute infection is associated with the tachyzoite form that spreads rapidly within the host. However, under stress conditions, some parasites can differentiate into cyst-forming bradyzoites, residing mainly in the central nervous system, retina and muscle. Because this latent form of the parasite is resistant to all currently available treatments, and is central to persistence and transmission of the parasite, specific therapeutic strategies targeting this developmental stage need to be found. T. gondii contains a plastid of endosymbiotic origin called the apicoplast, which is an appealing drug target because it is essential for tachyzoite viability and contains several key metabolic pathways that are largely absent from the mammalian host. Its function in bradyzoites, however, is unknown. Our objective was thus to study the contribution of the apicoplast to the viability and persistence of bradyzoites during chronic toxoplasmosis. We have used complementary strategies based on stage-specific promoters to generate conditional bradyzoite mutants of essential apicoplast genes. Our results show that specifically targeting the apicoplast in both in vitro or in vivo-differentiated bradyzoites leads to a loss of long-term bradyzoite viability, highlighting the importance of this organelle for this developmental stage. This validates the apicoplast as a potential area to look for therapeutic targets in bradyzoites, with the aim to interfere with this currently incurable parasite stage.

[Needle-embedding treatment for hemifacial spasm: research progress].

The articles regarding needle-embedding treatment for hemifacial spasm published before September 30, 2019 were searched from SinoMed, Wanfang, CNKI, VIP and PubMed database, and were analyzed and summarized from treatment methods, acupoint selection, stage differentiation and action mechanism. As a result, 45 Chinese articles were obtained. The needle-embedding treatment was divided into intradermal needling and acupoint thread-embedding; the top five acupoints were Sibai (ST 2), Taiyang (EX-HN 5), Dicang (ST 4), Jiache (ST 6) and spasm trigger points. The basic research of needle-embedding treatment for hemifacial spasm is weak, and the literature regarding stage differentiation is insufficient, which are in need of further study.

Needle-embedding treatment for hemifacial spasm: research progress / 中国针灸

The articles regarding needle-embedding treatment for hemifacial spasm published before September 30, 2019 were searched from SinoMed, Wanfang, CNKI, VIP and PubMed database, and were analyzed and summarized from treatment methods, acupoint selection, stage differentiation and action mechanism. As a result, 45 Chinese articles were obtained. The needle-embedding treatment was divided into intradermal needling and acupoint thread-embedding; the top five acupoints were Sibai (ST 2), Taiyang (EX-HN 5), Dicang (ST 4), Jiache (ST 6) and spasm trigger points. The basic research of needle-embedding treatment for hemifacial spasm is weak, and the literature regarding stage differentiation is insufficient, which are in need of further study.

Protein kinase A signaling during bidirectional axenic differentiation in Leishmania.

Parasitic protozoa of the genus Leishmania are obligatory intracellular parasites that cycle between the phagolysosome of mammalian macrophages, where they proliferate as intracellular amastigotes, and the midgut of female sand flies, where they proliferate as extracellular promastigotes. Shifting between the two environments induces signaling pathway-mediated developmental processes that enable adaptation to both host and vector. Developmentally regulated expression and phosphorylation of protein kinase A subunits in Leishmania and in Trypanosoma brucei point to an involvement of protein kinase A in parasite development. To assess this hypothesis in Leishmania donovani, we determined proteome-wide changes in phosphorylation of the conserved protein kinase A phosphorylation motifs RXXS and RXXT, using a phospho-specific antibody. Rapid dephosphorylation of these motifs was observed upon initiation of promastigote to amastigote differentiation in culture. No phosphorylated sites were detected in axenic amastigotes. To analyse the kinetics of (re)phosphorylation during axenic reverse differentiation from L. donovani amastigotes to promastigotes, we first established a map of this process with morphological and molecular markers. Upon initiation, the parasites rested for 6-12 h before proliferation of an asynchronous population resumed. After early changes in cell shape, the major changes in molecular marker expression and flagella biogenesis occurred between 24 and 33 h after initiation. RXXS/T re-phosphorylation and expression of the regulatory subunit PKAR1 correlated with promastigote maturation, indicating a promastigote-specific function of protein kinase A signaling. This is supported by the localization of PKAR1 to the flagellum, an organelle reduced to a remnant in amastigote forms. We conclude that a significant increase in protein kinase A-mediated phosphorylation is part of the ordered changes that characterise the amastigote to promastigote differentiation.

The DRBD13 RNA binding protein is involved in the insect-stage differentiation process of Trypanosoma brucei.

DRBD13 RNA-binding protein (RBP) regulates the abundance of AU-rich element (ARE)-containing transcripts in trypanosomes. Here we show that DRBD13 regulates RBP6, the developmentally critical protein in trypanosomatids. We also show DRBD13-specific regulation of transcripts encoding cell surface coat proteins including GPEET2, variable surface glycoprotein (VSG) and invariant surface glycoprotein (ISG). Accordingly, alteration in DRBD13 levels leads to changes in the target mRNA abundance and parasite morphology. The high consistency of the observed phenotype with known cell membrane exchanges that occur during progression of T. brucei through the insect stage of its life cycle suggests that DRBD13 is an important regulator in this largely unknown developmental process.