Blastocystis is a common and broadly distributed microbial eukaryote inhabiting the gut of humans and other animals. The genetic diversity of Blastocystis is extremely high comprising no less than 17 subtypes in mammals and birds. Nonetheless, little is known about the prevalence and distribution of Blastocystis subtypes colonising humans in Thailand. Molecular surveys of Blastocystis remain extremely limited and usually focus on the central, urban part of the country. To address this knowledge gap, we collected stool samples from a population of Thai adults (n = 178) residing in Chiang Rai Province. The barcoding region of the small subunit ribosomal RNA was employed to screen for Blastocystis and identify the subtype. Forty-one stool samples (23%) were identified as Blastocystis positive. Six of the nine subtypes that colonise humans were detected with subtype (ST) three being the most common (68%), followed by ST1 (17%) and ST7 (7%). Comparison of subtype prevalence across Thailand using all publicly available sequences showed that subtype distribution differs among geographic regions in the country. ST1 was most commonly encountered in the central region of Thailand, while ST3 dominated in the more rural north and northeast regions. ST2 was absent in the northeast, while ST7 was not found in the center. Thus, this study shows that ST prevalence and distribution differs not only among countries, but also among geographic regions within a country. Potential explanations for these observations are discussed herewith.
Breast cancer (BC) is the most common cancer in women. Although standard treatments are successful in patients with BC diagnosed at an early stage, an alternative treatment is required for patients with advanced-stage disease who do not respond to these treatments. The concept of using chemotherapy to sensitize cancer cells to become susceptible to immunotherapy was recently introduced and may be used as an alternative treatment for BC. The chemotherapeutic drug doxorubicin has been reported to sensitize cancer cells; however, the efficacy to sensitize the solid spheroids, in addition to its underlying mechanism regarding how doxorubicin sensitizes BC, has not previously been explored. In the present study, the effectiveness of a combined treatment of doxorubicin and natural killer-92 (NK-92) cells against BC in either 2D or 3D spheroid models, and its association with Fas receptor (FasR) expression, was demonstrated. The BC (MCF7) cell line expressing a higher level of FasR was more sensitive to NK-92 cell killing than the MDA-MB-231 cell line, which expressed a lower level of FasR. A sublethal dose of doxorubicin caused a significant improvement in NK cytotoxicity. Concordantly, a significant reduction in cell viability was observed in the doxorubicin-treated MCF7 spheroids. Notably, flow cytometric analysis revealed significantly increased FasR expression in the MCF7 cells, suggesting the underlying sensitization mechanism of doxorubicin in BC was related to the FasR upregulation. The present findings supported the use of combined doxorubicin and NK immunotherapy in BC treatment.
Abstractβ-thalassaemia is one of the most common genetic blood diseases worldwide with over 300 mutations in the HBB gene affecting red blood cell functions. Recently, advances in genome editing technology have provided a powerful tool for precise genetic correction. Generation of patient-derived induced pluripotent stem cells (iPSCs) followed by genetic correction of HBB mutations and differentiation into haematopoietic stem/progenitor cells (HSPCs) offers a potential therapy to cure the disease. However, the biggest challenge is to generate functional HSPCs that are capable of self-renewal and transplantable. In addition, functional analyses of iPSC-derived erythroid cells are hampered by poor erythroid expansion and incomplete erythroid differentiation. Previously, we generated an immortalised erythroid cell line (SiBBE) with unique properties, including unlimited expansion and the ability to differentiate into mature erythrocytes. In this study, we report a highly efficient genetic correction of HbE mutation in the SiBBE cells using the CRISPR/Cas9 system. The HbE-corrected clones restored β-globin production with reduced levels of HbE upon erythroid differentiation. Our approach provides a sustainable supply of corrected erythroid cells and represents a valuable model for validating the therapeutic efficacy of gene editing systems.
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