Folate receptor alpha targeted delivery of artemether to breast cancer cells with folate-decorated human serum albumin nanoparticles.

The pharmaceutical application of artemether (ARM) as an anticancer natural agent is hampered due to its poor solubility and bioavailability. In the present study, ARM was encapsulated in human serum albumin nanoparticles (HSA NPs) via desolvation method led to improvement of the water solubility by 50 folds. In further, folate-decorated ARM-HSA NPs (F-ARM-HSA NPs) were developed to enhance targeted delivery to folate receptor alpha (FRα)-overexpressing breast cancer cells. The hydrodynamic diameter and the zeta potential value of F-ARM-HSA NPs were 198 ± 11.22 nm and -23 ± 0.88 mV, respectively. Fluorescent microscopy demonstrated an enhanced cellular uptake of F-ARM-HSA NPs by high FRα-expressing MDA-MB-231 breast cancer cells compared to low FRα-expressing SK-BR-3 breast cancer cells. Cytotoxicity assay revealed a small significant difference between cytotoxicity effect of targeted and non-targeted NPs in SK-BR-3 cells. However, in MDA-MB-231 cells due to FRα-mediated endocytosis, the F-conjugated NPs had less inhibitory concentration (IC50) value (19.82 µg/mL) and higher cytotoxicity after 72 h compared to non-targeted ARM-HSA NPs. Flow cytometry analysis indicated a more potent drug-induced apoptosis rather than necrosis. The results suggest that our novel F-ARM-HSA NPs are likely to be recommended as a promising candidate for combination therapy of FRα-overexpressing breast cancer cells.

CAR-modified T-cell therapy for cancer: an updated review.

The use of chimeric antigen receptor (CAR)-modified T cells is a promising approach for cancer immunotherapy. These genetically modified receptors contain an antigen-binding moiety, a hinge region, a transmembrane domain, and an intracellular costimulatory domain resulting in T-cell activation subsequent to antigen binding. Optimal tumor removal through CAR-modified T cells requires suitable target antigen selection, co-stimulatory signaling domain, and the ability of CAR T cells to traffic, persist, and retain antitumor function after adoptive transfer. There are several elements which can improve antitumor function of CAR T cells, including signaling, conditioning chemotherapy and irradiation, tumor burden of the disease, T-cell phenotype, and supplementary cytokine usage. This review outlines four generations of CAR. The pre-clinical and clinical studies showed that this technique has a great potential for treatment of solid and hematological malignancies. The main purpose of the current review is to focus on the pre-clinical and clinical developments of CAR-based immunotherapy.

Designing novel and simple competitive internal amplification control for reliable PCR diagnosis of herpes simplex virus.

BACKGROUND: PCR is a molecular technique for herpes simplex virus (HSV) detection that can cause life-threatening infections such as encephalitis and keratitis. However, the main issues, false-negative results causing by PCR inhibitors, of this technique that reduce PCR efficiency. To overcome this problem, a competitive internal amplification control (IAC) was constructed for conventional PCR using the PCR-cloning technique. OBJECTIVES: The purpose of this study is the design of competitive IAC for PCR diagnosis of HSV, which in fact is the main cause of keratitis and viral encephalitis in developed countries. MATERIALS AND METHODS: Composite primers for PCR amplification of Leishmania major kDNA (kinetoplast DNA) were designed and optimized to use as IAC-HSV. IAC-HSV amplified in a non-stringent condition, ligated into pTZ57R plasmid vector, and transformed into Escherichia coli JM207 and then cloned. Resulting IAC was used for 105 CSF and 78 keratitis specimens. RESULTS: PCR amplicons for HSV and IAC-HSV were 454-bp and 662-bp, respectively. Detection limit of IAC was determined as 1000 plasmids per PCR reaction. IAC sensitivity for HSV detection was determined as 1000 plasmids per PCR reaction. IAC sensitivity for HSV detection was 500 copies/mL of HSV DNA. Among all specimens, 7 inhibited specimens were detected. CONCLUSIONS: Indeed, using other DNA as an IAC is expected to detect false-negative results and amplification of the DNA is the key tool to examine the accuracy of amplification and detection steps. This internal amplification control is applicable for early reliable diagnosis of HSV in different loads of virus in different specimens.