C-Terminal Domain Deletion Enhances the Protective Activity of cpa/cpb Loaded Solid Lipid Nanoparticles against Leishmania major in BALB/c Mice

Cutaneous leishmaniasis (CL) is the most common form of leishmaniasis with an annual incidence of approximately 2 million cases and is endemic in 88 countries, including Iran. CL's continued spread, along with rather ineffectual treatments and drug-resistant variants emergence has increased the need for advanced preventive strategies. We studied Type II cysteine proteinase (CPA) and Type I (CPB) with its C-terminal extension (CTE) as cocktail DNA vaccine against murine and canine leishmaniasis. However, adjuvants' success in enhancing immune responses to selected antigens led us to refocus our vaccine development programs. Herein, we discuss cationic solid lipid nanoparticles' (cSLN) ability to improve vaccine-induced protective efficacy against CL and subsequent lesion size and parasite load reduction in BALB/c mice. For this work, we evaluated five different conventional as well as novel parasite detection techniques, i.e., footpad imaging, footpad flowcytometry and lymph node flowcytometry for disease progression assessments. Vaccination with cSLN-cpa/cpb-CTE formulation showed highest parasite inhibition at 3-month post vaccination. Immunized mice showed reduced IL-5 level and significant IFN-ã increase, compared to control groups. We think our study represents a potential future and a major step forward in vaccine development against leishmaniasis

Reduced ikk/nf-kb expression by nigella sativa extract in breast cancer

Background: Activation of IKK/NF-kB signaling pathway plays a critical role in inflammation-driven tumor progression. Several natural compounds able to inhibit the IKK/NF-kB activation pathway have been shown to either prevent cancer or inhibit cell growth. Extensive studies have been carried out on the Nigella sativa (N. sativa) by many researchers, and its pharmacological activities including anticancer, analgesic, and anti-inflammatory functions have been explored. This study investigated the effect of N. sativa extract on the mRNA level of NFk (p50, RelB) and IKK (IKKA, IKKB) to determine one of the anti-inflammatory mechanisms of N. sativa in breast cancer cells. Methods: In this experimental study, MCF7 cell line was treated with different concentrations of hydroalcoholic extracts of N. sativa (0, 200, 400,600,800 μg/mL) for 24, 48 and 72 h. Effects of the extract on cell viability and NFk (p50, RelB ) and IKK (IKKA, IKKB) gene expression were analyzed by MTT assay and real time PCR, respectively. Results: mRNA expression levels of NFk (p50, RelB) and IKK (IKKA, IKKB) in the treatment group were lower than the untreated (control) group. Fold difference (p50, RelB) of gene expressions in treatment groups were statistically significant (P =0.001 and P =0.003) and the fold difference of IKK (IKKA, IKKB) in the treatment groups was lower than that of the untreated groups (P=0.01 and P=0.001). Conclusion: One possible anti-inflammatory mechanism of N. sativa is associated with the reduction in mRNA levels of NFk (p50, RelB) and IKK (IKKA, IKKB) in breast cancer. © 2020, Shiraz University of Medical Sciences. All rights reserved

Curcumin loaded on graphene nanosheets induced cell death in mammospheres from MCF-7 and primary breast tumor cells

Elimination of tumor cells is still a therapeutic challenge for breast cancer (BC) in men and women. Mammospheres serve as valuable in vitro tools for evaluating tumor behavior and sensitivity to anticancer treatments. Graphene nanosheets with unique physicochemical properties have been considered as potential biomedical approaches for drug delivery, bioimaging, and therapy. Graphene oxide (GO) and graphene quantum dots (GQDs) are suitable nanocarriers for hydrophobic and low bioaccessible anti-tumor materials like curcumin. Despite extensive studies on the potential application of graphene nanosheets in medicine, our knowledge of how different cells function and respond to these nanoparticles remains limited. Here, we evaluated cell death in mammospheres from MCF-7 and primary tumor cells in response to curcumin loaded on graphene nanosheets. Mammospheres were exposed to graphene oxide-curcumin (GO-Cur) and graphene quantum dots-curcumin (GQDs-Cur), and the incidence of cell death was evaluated by Hoechst 33342/propidium iodide double staining and flow cytometry. Besides, the expression of miR-21, miR-29a, Bax, and Bcl-2 genes were assessed using RT-qPCR. We observed, GO, and GQDs had no cytotoxic effect on Kerman male breast cancer/71 (KMBC/71) and MCF-7 tumor cells, while curcumin induced death in more than 50 of tumor cells. GO-Cur and GQDs-Cur synergistically enhanced anti-tumor activity of curcumin. Moreover, GQDs-Cur induced cell death in almost all cells of KMBC/71 mammospheres (99; p < 0.0001). In contrast, GO-Cur induced cell death in only 21 of MCF-7 mammosphere cells (p < 0.0001). Also, the expression pattern of miR-21, miR-29a, and Bax/Bcl-2 ratio in KMBC/71 and MCF-7 mammospheres was different in response to GO-Cur and GQDs-Cur. Although KMBC/71 and MCF-7 tumor cells had similar clinical features and displayed similar responses to curcumin, more investigations are needed to clarify the detailed molecular mechanisms underlying observed differences in response to GO-Cur and GQDs-Cur. © 2021 IOP Publishing Ltd