Background: Humans may be accidently infected by larva stage of Toxocara canis and Toxocara cati nematodes through consumption of contaminated vegetables and food by embryonated eggs and geophagia. They cause visceral, ocular, and neurological syndromes detected through serological methods. Parasitic contamination and infectious diseases are the main cause of eosinophilia. Objectives: The current study aimed at investigating Toxocara canis seroprevalence in military personnel and their families with eosinophilia referred to a military hospital in Tehran, Iran, from 2015 to 2016. Methods: In the current cross sectional study, 179 patients (military personnel and their families) referred to a military hospital with eosinophilia > 5% in a two-year period (2015 -2016) were selected and after obtaining informed consent and filling out the questionnaires, anti-Toxocara canis IgG was detected in their sera using the enzyme-linked immunosorbent assay (ELISA) technique. Results: Seroprevalence of Toxocara canis infection was 11.7%. Males were the most infected individuals. After statistical analysis, a significant relationship between the level of education (P = 0.01) and history of pet contact (P = 0.02), and anti-T. canis IgG was found; nevertheless, it had no relationship with age, gender, occupation, and place of residence (P > 0.05). Conclusions: Prevalence of Toxocara antibody was relatively high, which can indicate susceptibility to Toxocara infection in military personnel due to military operations. Due to lack of clinical manifestations in some patients, survey of Toxocara infection in eosinophilia cases is recommended.
EZH2, as a histone methyltransferase, has been associated with cancer development and metastasis possibly through the regulation of microRNAs and cellular pathways such as EMT. In this study, the effect of EZH2 expression on miR-200c and important genes of the EMT pathway was investigated in esophageal squamous cell carcinoma (ESCC). Comparative qRT-PCR was used to examine EZH2 expression in ESCC lines (YM-1 and KYSE‐30) following the separately transfected silencing and ectopic expressional EZH2 vectors in ESCC. Subsequently, expression of miR-200c and EMT markers was also assessed using qRT-PCR, western blotting and immunocytochemistry. Underexpression of Mir200c was detected in YM-1 and KYSE-30 cells after EZH2 silencing, while its overexpression was observed after EZH2 induced expression. Following EZH2 silencing, downregulation of mesenchymal markers and upregulation of epithelial markers were detected in the ESCCs. Our results demonstrate that EZH2 regulates the expression of miR-200c and critical EMT genes, implying that overexpression of Zeb2, Fibronectin, N-cadherin, and Vimentin lead to a mesenchymal phenotype and morphology while underexpression of epithelial genes, enhance cell migration after enforced expression of EZH2 in ESCCs. EZH2 gene can be a beneficial treatment marker for patients with esophageal cancer through decrease invasiveness of the disease and efficient response to neoadjuvant therapy.
Human enterokinase light chain (hEKL) cDNA sequence was designed with the help of codon optimization towards Escherichia coli codon preference and ribosome binding site design and artificially synthesized with a thioredoxin fusion tag at the N-terminal and a five his-tag peptide at the C-terminal. The synthetic hEKL gene was cloned into the pET-15 expression vector and transferred into the three different expression strains of E. coli BL21(DE3), NiCo21, and SHuffle T7 Express. Different growth and induction conditions were studied using a statistical response surface methodology (RSM). Recombinant hEKL protein was expressed at high levels in soluble form with 0.71 mM IPTG after 4 h of induction at 25 °C. Autocatalytic process cleaved TRX tag with enterokinase recognition site by the impure hEKL and yielded the mature enzyme. The target protein was then purified to homogeneity (> 95%) by affinity chromatography. The activity of hEKL was comparable to the commercial enzyme. From 1 L culture, 80 mg pure active hEKL was obtained with the specific activity of 6.25 × 102 U/mg. Three main parameters that help us to produce the enzyme in the folded and active form are the type of strain, SHuffle T7 strain, TRX and histidine fusion tags, and growth conditions including the increase of OD of induction and IPTG concentration and the decrease of induction temperature.
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