Optimization of a recombinant BlaR-CTD protein formulation using the response surface methodology.

The sequence of a carboxy-terminal of the ß-lactam sensor-transducer protein (BlaR-CTD) from Bacillus licheniformis ATCC14580 was extracted from US7745193B2 patent and expressed in E. coli using pColdI vector as a soluble His-tag recombinant protein. In this study, several excipients were used to improve the stability of recombinant BlaR-CTD and obtain the optimal formulation for this protein using response surface methodology (RSM)/ Central Composite Design (CCD). Total protein concentration was measured by UV spectroscopy and the Bradford test. A total of 7 various factors were designed using four different excipients including Glycerol, Sucrose, Triton x-100, and Tween-20, and three different buffers like Tris, Borate, and PBS. By obtaining suitable excipients and buffer i.e. glycerol and sucrose, pH ranging from 7 to 9 were evaluated. The pH 7.62, glycerol 15.35%, and sucrose 152.52 mM were determined as the most suitable for improving the thermal stability of recombinant BlaR-CTD.

Anticancer, antioxidant, and antibacterial effects of nanoemulsion of Origanum majorana essential oil.

Background and Objectives: This study aimed to develop a natural nanoemulsion with antibacterial and anticancer properties. Materials and Methods: The chemical composition of the Origanum majorana essential oil was investigated using GC-MS analysis. Besides, the successful loading of the essential oil in the nanoemulsion was confirmed using ATR-FTIR analysis. Moreover, nanoemulsion's anticancer, antioxidant, and antibacterial activities were investigated. Results: Terpinen-4-o1 (46.90%) was identified as the major compound in the essential oil. The nanoemulsion with a 149 ± 5 nm droplet size and zeta potential of -11 ± 1 mV was prepared. The cytotoxic effect of the nanoemulsion against A-375 human melanoma cells (IC50 = 139 µg/mL) showed significantly more potency than A-549 human lung cancer cells (IC50 = 318 µg/mL). Interestingly, growth of Staphylococcus aureus (Gram-positive) and E. coli (Gram-negative) bacteria after treatment with 4800 µg/mL of nanoemulsion were obtained at 12 ± 2 and 6 ± 1%, respectively. However, the IC50 value of nanoemulsion against E. coli (580 µg/mL) was not significantly different (P > 0.05) from S. aureus (611 µg/mL). Conclusion: A straightforward preparation method, high stability, and multi-biological effects are the main advantages of the prepared nanoemulsion. Therefore it could be considered for further investigation in vivo studies or complementary medicine.

Antioxidative, anticancer, and antibacterial activities of a nanogel containing Mentha spicata L. essential oil and electrospun nanofibers of polycaprolactone-hydroxypropyl methylcellulose.

BACKGROUND: As the largest organ, the skin has been frequently affected by trauma, chemical materials, toxins, bacterial pathogens, and free radicals. Recently, many attempts have been made to develop natural nanogels that, besides hydrating the skin, could also be used as antioxidant or antibacterial agents. METHODS: In this study, the chemical composition of the Mentha spicata essential oil was first investigated using GC-MS analysis. Its nanoemulsion-based nanogel was then investigated; successful loading of the essential oil in the nanogel was confirmed using FTIR analysis. Besides, nanogel's antioxidative, anticancer, and antibacterial activities were investigated. RESULTS: Carvone (37.1%), limonene (28.5%), borneol (3.9%), ß-pinene (3.3%), and pulegone (3.3%) were identified as five major compounds in the essential oil. By adding carboxymethylcellulose (3.5% w/v) to the optimal nanoemulsion containing the essential oil (droplet size of 196 ± 8 nm), it was gelified. The viscosity was fully fitted with a common non-Newtonian viscosity regression, the Carreau-Yasuda model. The antioxidant effect of the nanogel was significantly more potent than the essential oil (P < 0.001) at all examined concentrations (62.5-1000 µg/mL). Furthermore, the potency of the nanogel with an IC50 value of 55.0 µg/mL was substantially more (P < 0.001) than the essential oil (997.4 µg/mL). Also, the growth of Staphylococcus aureus and Escherichia coli after treatment with 1000 µg/mL nanogel was about 50% decreased compared to the control group. Besides, the prepared electrospun polycaprolactone-hydroxypropyl methylcellulose nanofibers mat with no cytotoxic, antioxidant, or antibacterial effects was proposed as lesion dressing after treatment with the nanogel. High potency, natural ingredients, and straightforward preparation are advantages of the prepared nanogel. Therefore, it could be considered for further consideration in vivo studies.

Nanogels Containing Foeniculum vulgare Mill. and Mentha piperita L. Essential Oils: Mosquitoes' Repellent Activity and Antibacterial Effect.

Foeniculum vulgare Mill. and Mentha piperita L. are two common medicinally important plants with a wide range of biological activities such as insecticide and antibacterial effects. In this study, the chemical composition of their essential oils was investigated using GC-MS analysis. After that, their nanoemulsions were prepared; optimum samples with droplet sizes of 74 ± 7 and 136 ± 5 nm were gelified. The viscosity of the prepared nanogels and the successful loading of the essential oil in them were investigated. The efficacy of the nanogel containing M. piperita essential oil as a repellent and antibacterial agent was more potent than the nanogel containing F. vulgare essential oil. Its completely protected time against Anopheles stephensi, the main malaria mosquito vector, was 120 ± 8 min. Moreover, the growth of Escherichia coli and Staphylococcus aureus after treatment with 5000 µg/mL of nanogel containing M. piperita essential oil was reduced by 100 and 65%, respectively. Considering natural constituents, a straightforward preparation method, and high efficacy, the nanogel containing M. piperita essential oil could be introduced for further investigation against other mosquitoes and bacterial species.

Cancer Stem Cells and Chemoresistance in Glioblastoma Multiform: A Review Article.

Glioblastomamultiforme (GBM) is the most common malignant and aggressive primary tumor of the brain in adults and characterized by a heterogeneous population of cells that are genetically unstable, highly infiltrative, angiogenic, and resistant to chemotherapy. Considerable efforts being devoted to identifying the molecular basis of resistance in GBM and exploring novel therapeutic targets that may improve overall survival. Several independent DNA repair mechanisms that normally safeguard genome integrity can facilitate drug resistance and cancer cell survival by removing chemotherapy- induced adducts. The recent data suggest that the most important mechanism of resistance to alkylating agents is the DNA repair enzyme O6-methylguanine methyltransferase (MGMT). Although, the treatment failure is a result of a number of causes, but currently, it has been demonstrated that a highly tumorigenic subpopulation of cancer cells called glioblastoma stem cells (GSCs) display relative resistance to radiation and chemotherapy. In fact, GBM stem cells express high levels of MGMT and this may account for GBM resistance following chemotherapy. GSCs also contribute to tumor growth through the stimulation of angiogenesis, which has been shown to be a useful therapeutic target in the treatment of recurrent or progressive malignant gliomas. In this review, we summarize the chemoresistance mechanisms of GBMs (to alkylating agents), with a special focus on the role of cancer stem cells.

An overview of the role of cancer stem cells in spine tumors with a special focus on chordoma.

Primary malignant tumors of the spine are relatively rare, less than 5% of all spinal column tumors. However, these lesions are often among the most difficult to treat and encompass challenging pathologies such as chordoma and a variety of invasive sarcomas. The mechanisms of tumor recurrence after surgical intervention, as well as resistance to radiation and chemotherapy, remain a pervasive and costly problem. Recent evidence has emerged supporting the hypothesis that solid tumors contain a sub-population of cancer cells that possess characteristics normally associated with stem cells. Particularly, the potential for long-term proliferation appears to be restricted to subpopulations of cancer stem cells (CSCs) functionally defined by their capacity to self-renew and give rise to differentiated cells that phenotypically recapitulate the original tumor, thereby causing relapse and patient death. These cancer stem cells present a unique opportunity to better understand the biology of solid tumors in general, as well as targets for future therapeutics. The general objective of the current study is to discuss the fundamental concepts for understanding the role of CSCs with respect to chemoresistance, radioresistance, special cell surface markers, cancer recurrence and metastasis in tumors of the osseous spine. This discussion is followed by a specific review of what is known about the role of CSCs in chordoma, the most common primary malignant osseous tumor of the spine.