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DNA vaccines with their extraordinary properties are the best choice as vectors for subunit vaccines but are not in compliance with safety regulations, mainly because of the antibiotic resistance genes on their backbone. New generations of plasmids with minimum bacterial backbones are now developed as promising alternatives to pass the safety rules and be replaced for conventional plasmids. Here we have compared the nanoplasmid (with RNA-out selection system and professional HTLV-1 containing promoter) and the conventionally used pcDNA plasmid, as regards the transfection efficiency. The EGFP gene was cloned in both pcDNA-3.1+ and NTC9385R-MSC and transfected into COS-7 cells for expression evaluation by flow cytometry. Meanwhile, qPCR was used to analyze the EGFP mRNA copy numbers. It was concluded that the nanoplasmid, with its extraordinary properties, can be a tempting alternative to conventional pcDNA in equal or equimolar concentrations for vaccine design. These promising results can put DNA vaccines back into focus, especially regarding diseases controlled by robust cellular immune responses. [ABSTRACT FROM AUTHOR]

Protein homeostasis is an equilibrium of paramount importance that maintains cellular performance by preserving an efficient proteome. This equilibrium avoids the accumulation of potentially toxic proteins, which could lead to cellular stress and death. While the regulators of proteostasis are the machineries controlling protein production, folding and degradation, several other factors can influence this process. Here, we have considered two factors influencing protein turnover: the subcellular localization of a protein and its functional state. For this purpose, we used an imaging approach based on the pulse-labeling of 17 representative SNAP-tag constructs for measuring protein lifetimes. With this approach, we obtained precise measurements of protein turnover rates in several subcellular compartments. We also tested a selection of mutants modulating the function of three extensively studied proteins, the Ca2+ sensor calmodulin, the small GTPase Rab5a and the brain creatine kinase (CKB). Finally, we followed up on the increased lifetime observed for the constitutively active Rab5a (Q79L), and we found that its stabilization correlates with enlarged endosomes and increased interaction with membranes. Overall, our data reveal that both changes in protein localization and functional state are key modulators of protein turnover, and protein lifetime fluctuations can be considered to infer changes in cellular behavior. [ABSTRACT FROM AUTHOR]

Aims: We investigated the protective effect of chitin micro‐particle (CMP) as an adjuvant against Leishmania infection in BALB/c mice. Methods: Mice were immunized subcutaneously with soluble Leishmania antigen (SLA) plus CMP (100 µg SLA + 100 µg CMP/100 µL) as the test group. Three weeks after the last immunization, test and control groups were infected by Leishmania major (L major). Eight weeks post‐infection, evaluation of parasites load in lymph nodes was performed using limiting dilution assay. Then, the spleen cell cytokine response (TNF‐α, IFN‐γ, IL‐4, IL‐10, IL‐17 and IL‐27) to SLA among vaccinated and nonvaccinated groups was investigated using ELISA. Serum levels of IgG1 and IgG2a were measured as well. Results: The SLA plus CMP group demonstrated the protection. The responses included reduced lesion formation and lower parasite load. Also, in comparison with control group higher levels of IFN‐γ and, IL‐10 in the culture of spleen cells, and lower levels of IgG1 in sera were seen in SLA plus CMP group. Conclusion: The data supported the possibility of using CMP as a suitable adjuvant in Leishmania vaccination. [ABSTRACT FROM AUTHOR]

Mitochondria contain their own gene expression machinery, which synthesizes core subunits of the oxidative phosphorylation system. Monitoring mitochondrial translation within spatial compartments of cells is difficult. Here we describe a method to visualize mitochondrial translation within defined parts of cells, using a click chemistry approach. This method can be applied to different cell types such as neurons and allows detection of newly synthesized mitochondrial proteins in spatial resolution using microscopy techniques. Furthermore, using click chemistry, mitochondrial translation can also be monitored by standard SDS-PAGE. The described method avenues the analysis of newly synthesized mitochondrial encoded proteins in the cellular context, by avoiding the usage of radioactive components., (Copyright © 2024. Published by Elsevier Inc.)

Background: Considering the incidence and prevalence rates of gastric cancer in Mazandaran Province of Iran, this research was performed to evaluate the efficacy and safety of olanzapine in symptom relief and quality of life (QOL) improvement of gastric patients receiving chemotherapy., Materials and Methods: This clinical trial was conducted on thirty new cases of gastric cancer patients whose treatment protocol was planned on chemotherapy and were allocated into two groups by simple random sampling. Intervention group (15 patients) received olanzapine tablets (2.5-10 mg/day) a day before the beginning of chemotherapy; in the 1 st day of chemotherapy to 8 weeks after chemotherapy, besides the routine treatment regimens. The control group received only the routine treatment regimens. The patients were followed for 8 weeks after intervention. All of the patients were assessed with Hospital Anxiety and Depression Scale (HADS) and WHO-QOL-BREF questionnaires; further, Rhodes index was used to evaluate nausea and vomiting (N/V) status., Results: All the recruited patients continued the allocated interventions (no lost to follow-up). N/V decreased in the case group, but the difference was not statistically significant ( P = 0.438). The patients' appetite and body mass index increased ( P = 0.006). Anxiety and depression subscales of HADS had significant differences between the two groups ( P < 0.001) in the 4 th and 8 th week after treatment. Among the different subdomains of QOL, only physical health improved significantly after intervention ( P < 0.05), but no significant difference was observed in other subdomains and also total QOL score ( P > 0.05). No significant increase was observed in fasting and 2-h postprandial blood glucose and lipid profile ( P > 0.05)., Conclusion: Olanzapine can be considered as an effective drug to increase appetite and decrease anxiety and depression in patients with gastric cancer., Competing Interests: There are no conflicts of interest.