In silico design of a multi-epitope vaccine against HPV16/18.

BACKGROUND: Cervical cancer is the fourth most common cancer affecting women and is caused by human Papillomavirus (HPV) infections that are sexually transmitted. There are currently commercially available prophylactic vaccines that have been shown to protect vaccinated individuals against HPV infections, however, these vaccines have no therapeutic effects for those who are previously infected with the virus. The current study's aim was to use immunoinformatics to develop a multi-epitope vaccine with therapeutic potential against cervical cancer. RESULTS: In this study, T-cell epitopes from E5 and E7 proteins of HPV16/18 were predicted. These epitopes were evaluated and chosen based on their antigenicity, allergenicity, toxicity, and induction of IFN-γ production (only in helper T lymphocytes). Then, the selected epitopes were sequentially linked by appropriate linkers. In addition, a C-terminal fragment of Mycobacterium tuberculosis heat shock protein 70 (HSP70) was used as an adjuvant for the vaccine construct. The physicochemical parameters of the vaccine construct were acceptable. Furthermore, the vaccine was soluble, highly antigenic, and non-allergenic. The vaccine's 3D model was predicted, and the structural improvement after refinement was confirmed using the Ramachandran plot and ProSA-web. The vaccine's B-cell epitopes were predicted. Molecular docking analysis showed that the vaccine's refined 3D model had a strong interaction with the Toll-like receptor 4. The structural stability of the vaccine construct was confirmed by molecular dynamics simulation. Codon adaptation was performed in order to achieve efficient vaccine expression in Escherichia coli strain K12 (E. coli). Subsequently, in silico cloning of the multi-epitope vaccine was conducted into pET-28a ( +) expression vector. CONCLUSIONS: According to the results of bioinformatics analyses, the multi-epitope vaccine is structurally stable, as well as a non-allergic and non-toxic antigen. However, in vitro and in vivo studies are needed to validate the vaccine's efficacy and safety. If satisfactory results are obtained from in vitro and in vivo studies, the vaccine designed in this study may be effective as a therapeutic vaccine against cervical cancer.

Protocol for isolating extracellular vesicles from pancreatic cancer cells for liver metastatic niche research.

Extracellular vesicles (EVs) are secreted, cell-derived, membrane-bound compartments implicated in various diseases for their ability to influence distant targets and as carriers of biomarkers. Here, we present a protocol for separating EVs from mammalian pancreatic cancer cells and their characterization using western blot and electron microscopy. We then demonstrate how they are utilized to affect tumor development in a murine model of metastatic pancreatic cancer including a method to quantify hepatic tumor burden in histologic samples. For complete details on the use and execution of this protocol, please refer to Dudgeon et al.1.

Prevalence of Gram-Negative Bacilli Isolated from the Equipment and Surfaces in Hospital Wards of Golestan Province, North of Iran.

Nosocomial infections are one of the most common causes of death in hospitals. This study aimed to determine the prevalence of gram-negative bacilli isolated from the equipment in hospital wards of the Golestan province, in the year 2015. In this cross-sectional study in 2015, 1980 samples from medical and nonmedical equipment and surfaces were collected from the wards of 13 teaching hospitals, in the Golestan province. Samples were inoculated into eosin methylene blue agar and blood agar culture media and isolated colonies were identified by standard biochemical tests. The obtained results were then analyzed using SPSS 22 software and χ2 test. Among 1980 isolated samples, 601 samples (30.35%) were infected with gram-negative bacilli while Enterobacter aerogenes (37.27%) was responsible for most of the contaminations. The highest rate of infection was observed in the intensive care unit (33.1%), and the highest level of contamination in the medical equipment was associated with laryngoscope and its blade (10.48%), as well as ECG sensor and its monitoring connector (6.65%). Meanwhile, phone (6.32%) and patients' beds and linen (5.15%) had the highest level of contamination in the nonmedical equipment. Considering the high rates of gram-negative bacilli contamination in the hospital wards of the Golestan province, thorough hand washing as the main action for disinfection and sterilizing the equipment, as well as performing periodic cultivation alongside the use of standard guidelines for prevention and control of nosocomial infections, are recommended to reduce the level of contamination.