The Inflammatory and Fibrotic Patterns of Hepatic Stellate Cells Following Coagulation Factors (VII or X)-Shielded Adenovirus Infection.

The role of coagulation factors on the inflammatory effect of adenovirus (Ad) is an unresolved question that was considered herein. Adenovirus-36(Ad36) and adenovector-5-GFP(Ad5-GFP) were prepared; then, they were loaded with VII or FX factors. The size/charge parameters and transduction efficiency were evaluated using fluorescent microscopy and Zetasizer, respectively. The Ad36-coagulation factor complexes were added on the stellate cells, LX-2. Thereafter, the expression levels of inflammatory and fibrotic genes including PKR, IL-1ß, TNF-α, TIMP-1, collagen, and TGF-ß were measured by qPCR and ELISA assays. The loading of FVII or FX factors not only increased the size/charge of Ad5-GFP but also enhanced the transduction rate up to 60% and 75%, respectively, compared to the controls (45%). The PKR expression analysis showed an upregulation following treatment with all Ad36 forms (P = 0.0152). The IL-1ß and TNF-α cytokines analyses demonstrated that the Ad36-FVII complex elicited the highest inflammatory response (P = 0.05). Similarly, the fibrosis-related expression analysis revealed a more inductive role of FVII when loaded on Ad36, compared to the FX factor. The findings suggested that adenovirus elicited the innate inflammatory and activation state in the hepatic stellate cell. In addition, adenovirus shielded by FVII exhibited more innate inflammation as well as activation of the stellate cells than the FX-loaded virus.

The Sero-Prevalence of Hepatitis B and C Viruses in Municipal Waste Collectors in Southwest of Iran.

Aims: The present study aimed to investigate the frequency of hepatitis B virus (HBV) and hepatitis C virus (HCV) serological markers among waste collectors in the municipality of Shiraz city, southwest of Iran, 2018. Settings and Design: In this cross-sectional study, a total of 385 waste collectors from all 10 districts of Shiraz city, southwest of Iran were enrolled. A questionnaire was used to gather occupational and demographic information as well as awareness about viral hepatitis. Methods and Material: Their blood samples were collected, and the sera were investigated for the presence of hepatitis B surface antigen (HBsAg), anti-HBs antibody, and anti-HCV antibody using enzyme-linked immunosorbent assay (ELISA). Results: All the participants were men with a mean age of 41 ± 8 years. Out of 385 participants, 6 (1.5%) subjects were positive for HBsAg, indicative of HBV infection. Moreover, 38 (9.9%) had a protective level of anti-HBs antibodies, while more than 90% had a low level of anti-HBs antibodies. All participants were negative for HCV antibodies. Conclusion: According to these findings, evaluation of HBsAg and anti-HBs levels is recommended in the healthy program of waste collector workers (WCWs). However, this occupation might not be a risk factor for the acquisition of HBV and especially HCV infections.

Normoxic HIF-1α Stabilization Caused by Local Inflammatory Factors and Its Consequences in Human Coronary Artery Endothelial Cells.

Knowledge about normoxic hypoxia-inducible factor (HIF)-1α stabilization is limited. We investigated normoxic HIF-1α stabilization and its consequences using live cell imaging, immunoblotting, Bio-Plex multiplex immunoassay, immunofluorescence staining, and barrier integrity assays. We demonstrate for the first time that IL-8 and M-CSF caused HIF-1α stabilization and translocation into the nucleus under normoxic conditions in both human coronary endothelial cells (HCAECs) and HIF-1α-mKate2-expressing HEK-293 cells. In line with the current literature, our data show significant normoxic HIF-1α stabilization caused by TNF-α, INF-γ, IL-1ß, and IGF-I in both cell lines, as well. Treatment with a cocktail consisting of TNF-α, INF-γ, and IL-1ß caused significantly stronger HIF-1α stabilization in comparison to single treatments. Interestingly, this cumulative effect was not observed during simultaneous treatment with IL-8, M-CSF, and IGF-I. Furthermore, we identified two different kinetics of HIF-1α stabilization under normoxic conditions. Our data demonstrate elevated protein levels of HIF-1α-related genes known to be involved in the development of atherosclerosis. Moreover, we demonstrate an endothelial barrier dysfunction in HCAECs upon our treatments and during normoxic HIF-1α stabilization comparable to that under hypoxia. This study expands the knowledge of normoxic HIF-1α stabilization and activation and its consequences on the endothelial secretome and barrier function. Our data imply an active role of HIF-1α in vivo in the vasculature in the absence of hypoxia.

The Evaluation of tLyP-1-Bound Mda-7/IL-24 Killing Activity on a Liver Tumor Cell Line.

Background: The melanoma differentiation-associated gene-7 (Mda-7)/interleukin-24 (IL-24) is a tumor killing cytokine, the bystander effect of which can be enhanced through tethering to tumor homing peptides (THPs). Materials and Methods: After fusing tLyP-1, RGR, and buforin as THPs to Mda-7/IL-24, enzyme-linked immunosorbent assay (ELISA) was used to determine the secretion potency of the recombinant proteins. The killing potency of plasmids expressing IL-24, IL-24.tLyP1, IL-24.RGR, and buf.IL-24 were assessed, using MTT, Annexin/PI staining assays, as well as measuring the expression level of GADD-153 and BCL2-associated X (BAX) on Huh-7 cells. Three-dimensional structural analysis and protein-receptor interaction were also evaluated by modeling. Results: The ELISA result showed that contrary to IL-24.RGR and buf.IL-24, IL-24.tLyP-1 retained the secretion potency, similar to the native form. The viability assessments showed that IL-24 and IL-24.tLyP-1 had the most growth suppressive effects in comparison with the control group (p < 0.0001). Furthermore, IL-24 and IL-24.tLyP-1 had the highest apoptotic activity and significant upregulatory effect on the GADD-153 and BAX genes (p < 0.0003). The modeling showed that peptide modifications left no detrimental effect on IL-24 attachment to the cognate receptor. Conclusion: IL-24 can tolerate tLyP-1 peptide modification by retaining its secretion potency. Tethering tLyP-1 to IL-24 can induce more apoptosis than its modified versions by RGR or buforin.

Evidence of Hantavirus circulation among municipal street sweepers, southwest of Iran.

Hantaviruses are rodent-borne zoonosis pathogens that cause hemorrhagic fever with renal syndrome (HFRS) and Hantavirus cardiopulmonary syndrome (HCPS) in humans. Rodents spread the virus via their excretions. The outbreak of Hantaviruses pose a significant public health problem. The epidemiology and history of Hantaviruses in Iran is not clear and regardless of the data from the few available studies, little is known about its epidemiology in this country. Herein, we discuss the prevalence of IgG antibody against Hantavirus serotypes in 385 street sweepers from southwest of Iran. Serum samples were investigated, using Hantavirus Pool 1 "Eurasia" IgG kit and Pool 2 "America" ELISA IgG kit (Euroimmun, Germany) to detect IgG antibodies against Old and New World Hantaviruses. The results showed a specific IgG antibody in two samples (0.5%). Both of seropositive cases had specific IgG antibody against Old World Hantaviruses. The data of the current study along with the previous data, indicate the circulation of Hantaviruses in Iran. Hence, the risk of Hantavirus infection in high-risk groups should be considered as a serious health issue.

The superior role of coagulation factor FX over FVII in adenoviral-mediated innate immune induction of the hepatocyte: an in vitro experiment.

AIM OF THE STUDY: To better understanding the contribution of coagulation factors to the extent of adenovirus-mediated innate toxicity on the hepatocyte. MATERIAL AND METHODS: Adenovirus-36 (AD) and adenovector type 5-GFP (Ad5-GFP) were propagated and titered; then, they were loaded with coagulation factors VII or X. The complex of adenovirus with coagulation factor VII and X were for size and charge parameters. After adding AD-VII and AD-X complexes, the expression levels of innate inflammatory genes including protein kinase R (PKR), interleukin (IL)-1ß, IL-8 and IL-18 were measured by Real-time PCR on a hepatocellular carcinoma cell line, HepG2. RESULTS: The loading of coagulation factors VII and X on Ad5-GFP enhanced the transduction rate up to 50% and 60% (p < 0.05), respectively, compared to the adenovector alone (30%) (p < 0.05). The formation of the coagulation factor-virus complex leads to multimodal size distribution with an increase in average hydrodynamic size and absolute zeta potential. The qPCR results showed that PKR expression increased significantly after treatment with all adenoviruses. These findings also showed that AD had a significant (p = 0.0152) inflammatory impact on Hep-G2. However, AD which was loaded with FX (AD-X) exhibited the most inflammatory effect (p = 0.0164). Significantly, the expression of IL-1ß (p = 0.0041), IL-8 (p = 0.0107) and IL-18 (p = 0.0193) were also enhanced following FX loading. On the other hand, the AD-VII complex showed the least effect of innate immune induction when compared to the negative control (p < 0.05). CONCLUSIONS: The loading of coagulation factors, particularly FX, could enhance the transduction efficiency of Ad5-GFP. Furthermore, adenovirus loaded with FX exhibited more innate toxicity on the hepatocytes, while it was not the case for FVII.