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Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of purified OMVs indicated protein profiles within the range of 34-52 kDa. Furthermore, transmission electron microscopy demonstrated the spherical shaped vesicles of 50-200 nm. The results of ELISA showed significant titers of systemic and mucosal immune anti-OMV IgGs in immunized BALB/c mice with different vaccine regimens. Additionally, a notable increase in the FA ratio was demonstrated in this study. The obtained results of the present study revealed that the WC-OMV combination of local strain can induce a high level of antibody response indicating more protection than OMV or WC separately. Moreover, it can be considered an effective immunogen against V. cholerae.Tuberculin skin test, also known as the tuberculin or purified protein derivative (PPD) test, is an extensively applied diagnostic test for the detection of primary infection with Mycobacterium tuberculosis (Mtb). The production of PPD is accompanied by some difficulties that require a series of modifications in the production and purification processes. The present study aimed to determine the facilitation level of the manufacturing process by modifying evaluation methods for the production of PPD tuberculin. Mtb strains were cultured in Lowenstein-Jensen media, and the cultured strains were inoculated into the Dorset-Henley liquid medium by the biphasic medium of potato-Dorset-Henley. After incubation, flasks containing cultured strain were selected for bacterial inactivation, and the optimal gamma radiation dose(s) was determined. Tuberculoproteins were precipitated by ammonium sulfate (AS) and Trichloroacetic acid (TCA). Protein concentration was determined using the Bradford and Kjeldahl protein assay mewas more efficiently carried out by the Kjeldahl method, compared to the Bradford method. Finally, the results of the present study demonstrated that in addition to the novel approach of gamma irradiation, optimum methods are efficient and applicable in the production of PPD tuberculin.The changes in temperature levels can potentially affect the toxins in terms of stability and immunological properties via alteration of their structures. Diphtheria Toxin (DT) is highly considered by scientists since its mechanism of action is similar to those of most bacterial toxins, such as botulinum, tetanus, and anthrax. The protection of conformational B-cell epitopes is critically important in the process of diphtheria vaccine production. This study aimed to evaluate the conformational changes of the DT structure at three different temperature levels (27˚C, 37˚C, and 47˚C) using molecular dynamic simulations. Secondary structures were analyzed in YASARA software. According to the results, significant decreases were observed in percentages of the β-sheets, turns, and the helices of the DT structure at 47˚C in comparison with those at 27˚C and 37˚C. Furthermore, the tertiary structure of the DT was compared at different temperatures using the contact map. Accordingly, the results showed that the root-mean-square deviation of the DT structure increased upon temperature rising. In addition, amino acids D68, G128, G171, C186, and K534-S535 at 27˚C and 37˚C, as well as amino acids G26, P38, S291, T267, H384, A356, and V518 at 47˚C showed higher root mean square fluctuation values. The finding demonstrated that the stability of the DT structure decreased at high temperature (47˚C). The solvent-accessible surface area diagram showed that the hydrophobicity of the DT structure increased via temperature rising, and the amino acid residues belonging to B-cell epitopes extended through increasing temperature. However, B-cell epitopes belonging to the junction region of chains A and B were only present at 37˚C. The results of this study are expected to be applicable for determining a suitable temperature level for the production process of the diphtheria vaccine.Toxoplasmosis is a widespread parasitic disease caused by a protozoan parasite Toxoplasma gondii. Currently, nanotechnology has been used for the diagnosis of many infectious diseases. It could be due to the fact that nanoparticles play an important role in accurate and fast diagnosis. The purpose of this study was to design a Nano-enzyme linked immunosorbent assay (Nano-ELISA) kit using excreted/secreted (E/S) antigens to have higher sensitivity and specificity than those reported for the designed enzyme-linked immunosorbent assay (ELISA) kit for the diagnosis of Toxoplasmosis in mice. Firstly, the serum samples were collected from 15 infected mice with T. gondii and 15 healthy ones. Then, E/S antigens were separated from parasite tachyzoites and used for designing an ELISA kit. In addition, the mice sera were evaluated using the designed ELISA kit. Finally, the serum samples were assessed by Nano-ELISA kits designed with E/S antigen and conjugate of gold nanoparticles. The obtained results of the present study showed that the sensitivity and specificity of the designed ELISA kit were reported as 80% and 86.66%, respectively, that both improved to 93.33% in these sera with the designed Nano-ELISA kit. This finding revealed the significant improvement of sensitivity and specificity using gold nanoparticles in designing the ELISA kit. Furthermore, according to the literature, the use of E/S antigens in designing recognizable ELISA kits has been always highlighted considering the presence of numerous antigens in T. gondii. The results of this study revealed that the use of E/S antigens in the preparation of an ELISA kit was very effective. This is very important, especially in the lower titers of antibody requiring a more accurate diagnosis. On the other hand, the Nano-ELISA method designed with E/S antigens can be more sensitive and specific than ELISA for the diagnosis of Toxoplasmosis and can be the basis for further studies in this regard.Pain occurs in a wide range of diseases and long-term conditions associated with ageing and can affect every aspect of an individual's life, reducing their ability to recover, their independence and their quality of life. U0126 order The assessment of pain is an important aspect of nurses' role and requires them to obtain detailed information on how the older person experiences pain and how pain is affecting their life. However, there are many challenges to effective pain assessment in older people, including challenges concerning communication and cognition. This article provides an overview of pain assessment in older people, particularly those living with dementia, based on the author's expertise and on relevant literature, notably the recently revised UK national guidelines on the assessment of pain in older people.