ABSTRACT
This review compared coronavirus disease 2019 (COVID-19) laboratory findings, comorbidities, and clinical outcomes in patients from the general population versus medical staff to aid diagnosis of COVID-19 in a more timely, efficient, and accurate way. Electronic databases were searched up to 23rdMarch, 2020. The initial search yielded 6,527 studies. Following screening, 24 studies were included [18 studies (11,564 cases) of confirmed COVID-19 cases in the general public, and 6 studies (394 cases) in medical staff] in this review. Significant differences were observed in white blood cell counts (p < 0.001), lymphocyte counts (p < 0.001), platelet counts (p = 0.04), procalcitonin levels (p < 0.001), lactate dehydrogenase levels (p < 0.001), and creatinine levels (p = 0.03) when comparing infected medical staff with the general public. The mortality rate was higher in the general population than in medical staff (8% versus 2%). This review showed that during the early stages of COVID-19, laboratory findings alone may not be significant predictors of infection and may just accompany increasing C-reactive protein levels, erythrocyte sedimentation rates, and lactate dehydrogenase levels. In the symptomatic stage, the lymphocyte and platelet counts tended to decrease. Elevated D-dimer fibrin degradation product was associated with poor prognosis.
ABSTRACT
Background: Leishmania major LmSTI1 is a conserved protein among different species of leishmania, and expressed in both amastigote and promastigote forms of L. major life cycle. It has previously been expressed in bacterial systems
Materials and Methods: To express LmSTI1 in the methylotrophic yeast Pichia pastoris [P. pastoris], the shuttle vector pPICZA containing gene lmsti1 was constructed under the control of the AOX1 promoter. The recombinant vector was electro-transformed into P. pastoris, and induced by 0.5% methanol in the buffered medium. The expression of the LmSTI1 protein was visualized in the total soluble protein of P. pastoris by 12% SDS-PAGE, and further confirmed by Western blotting with L.major-infected mouse sera and HRP-conjugated goat anti-mouse IgG as the first and secondary antibodies, respectively
Results: The expression level was 0.2% of total soluble proteins
Conclusion: It might be possible to use this formulation as a whole yeast candidate vaccine against cutaneous leishmanization
ABSTRACT
Background: Recent researches have demonstrated that human T-cell immunoglobulin mucin 1 [TIM-1] glycoprotein plays important roles in regulation of autoimmune and allergic diseases, as well as in tumor immunity and response to viral infections. Therefore, targeting TIM-1 could be a potential therapeutic approach against such diseases
Objectives: In this study, we aimed to express TIM-1 protein on Human Embryonic kidney [HEK] 293T cell line in order to have an available source of the TIM-1 antigen
Materials and Methods: The cDNA was synthesized after RNA extraction from peripheral blood mononuclear cells [PBMC] and TIM-1 cDNA was amplified by PCR with specific primers. The PCR product was cloned in pcDNA[TM]3.1/Hygro [+] and transformed in Escherichia coli TOP 10 F'. After cloning, authenticity of DNA sequence was checked and expressed in HEK 293T cells. Finally, expression of TIM-1 was analyzed by flow cytometry and real-time PCR
Results: The result of DNA sequencing demonstrated correctness of TIM-1 DNA sequence. The flow cytometry results indicated that TIM-1 was expressed in about 90% of transfected HEK 293T cells. The real-time PCR analysis showed TIM-1 mRNA expression increased 195-fold in transfected cells compared with un-transfected cells
Conclusions: Findings of present study demonstrated the successful cloning and expression of TIM-1 on HEK 293T cells. These cells could be used as an immunogenic source for production of specific monoclonal antibodies, nanobodies and aptamers against human TIM-1
ABSTRACT
The C-terminal region of the merozoite surface protein 1 (MSP-1) of Plasmodium falciparum is a strong vaccine candidate as it is associated with immunity to the parasite. This corresponds approximately to the conserved 17th block of the gene and is composed of two EGF- like domains. These domains exhibit only four single amino acid substitutions which show several potential variants in this region of the gene. As the variations might be important for a regional vaccine design, a study was carried out to determine the variations present in P. falciparum isolates from southern Iran. Besides the usual E-T-S-R-L and the Q-K-N-G-F types, we found Q-T-S-R-L, E-K-N-G-F, E-T-S-G-L, Z-T-S-G-L and Z-T-S-R-L types, where Z was E or Q signifying the presence of mixed clones in single isolates.
ABSTRACT
Expression of eukaryotic proteins in E. coli often results in their aggregation. Proper folding and solubility of therapeutical proteins are the pre-requisite for their bioactivity. This is not achieved in cytoplasmic expression in E. coli because of the absence of disulfide bonds formation. A novel expression/secretion vector was constructed which exploited beta -lactamase signal sequence to translocate processed and soluble proteins into the periplasm of cells. Secretion of model proteins, beta -lactamase and human Epidermal Growth Factor [hEGF] in M15/pSB and M15/pSE systems respectively, was confirmed by SDS-PAGE analysis and bioactivity assay. Secreted hEGF was found to be identical to authentic protein, in size, N-terminal amino acid sequence, biological activity and Western-blotting. The radioimmunoassay revealed 10-fold-higher level of hEGF expression in M15/pSE secretion system compared to that of cytoplasmic expression of the protein. The properly processed and in vivo folded hEGF demonstrated high solubility and bioactivity. The data obtained, evidenced in favor of M15/pSE expression system, which might be suitable to produce the small eukaryotic disulfide-bonded proteins for therapeutical applications or structural studies