HEV is the most causative agent of acute viral hepatitis globally. HEV causes acute, chronic, and extrahepatic manifestations. Chronic HEV infection develops in immunocompromised patients such as organ transplant patients, HIV-infected patients, and leukemic patients. The source of chronic HEV infection is not known. Also, the source of extrahepatic manifestations associated with HEV infection is still unclear. Hepatotropic viruses such as HCV and HBV replicate in peripheral blood mononuclear cells (PBMCs) and these cells become a source of chronic reactivation of the infections in allograft organ transplant patients. Herein, we reported that PBMCs and bone marrow-derived macrophages (BMDMs), isolated from healthy donors (n = 3), are susceptible to HEV in vitro. Human monocytes (HMOs), human macrophages (HMACs), and human BMDMs were challenged with HEV-1 and HEV-3 viruses. HEV RNA was measured by qPCR, the marker of the intermediate replicative form (ds-RNA) was assessed by immunofluorescence, and HEV capsid protein was assessed by flow cytometry and ELISA. HEV infection was successfully established in primary HMOs, HMACs, and human BMDMs, but not in the corresponding cells of murine origin. Intermediate replicative form (ds RNA) was detected in HMOs and HMACs challenged with HEV. The HEV load was increased over time, and the HEV capsid protein was detected intracellularly in the HEV-infected cells and accumulated extracellularly over time, confirming that HEV completes the life cycle inside these cells. The HEV particles produced from the infected BMDMs were infectious to naive HMOs in vitro. The HEV viral load was comparable in HEV-1- and HEV-3-infected cells, but HEV-1 induced more inflammatory responses. In conclusion, HMOs, HMACs, and human BMDMs are permissive to HEV infection and these cells could be the source of chronic and recurrent infection, especially in immunocompromised patients. Replication of HEV in human BMDMs could be related to hematological disorders associated with extrahepatic manifestations.

Hepatitis E virus (HEV) infection is endemic in developing and developed countries. HEV was reported to be excreted in the milk of ruminants, raising the possibility of transmission of HEV infection through the ingestion of contaminated milk. Therefore, the detection of HEV markers in milk samples becomes pivotal. However, milk includes inhibitory components that affect HEV detection assays. Previously it was reported that dilution of milk matrix improves the performance of HEV molecular assay, however, the dilution of milk samples is not the best strategy especially when the contaminated milk sample has a low HEV load. Therefore, the objective of this study is to compare the effect of extraction procedures on the efficiency of HEV RNA detection in undiluted milk samples. In addition, we assessed the effect of the removal of milk components such as fats and casein on the performance of the molecular and serological assays of HEV. Phosphate buffered saline (PBS) and different milk matrices (such as whole milk, skim milk, and milk serum) were inoculated with different HEV inoculums and subjected to two different extraction procedures. Method A includes manual extraction using spin column-based extraction, while method B includes silica-based automated extraction. Method A was more sensitive than method B in the whole milk and skim milk matrices with a LoD_95% of 300 IU/mL, and virus recovery yield of 47%. While the sensitivity and performance of method B were significantly improved using the milk serum matrix, with LoD_95% of 96 IU/mL. Interestingly, retesting HEV positive milk samples using the high sensitivity assay based on method B extraction and milk serum matrix increased the HEV RNA detection rate to 2-fold. Additionally, the performance of HEV serological assays such as anti-HEV IgG and HEV Ag in the milk samples was improved after the removal of the fat globules from the milk matrix. In conclusion, HEV RNA assay is affected by the components of milk and the extraction procedure. Removal of inhibitory substances, such as fat and casein from the milk sample increased the performance of HEV molecular and serological assays which will be suitable for the low load HEV milk with no further dilutions.

Background: Atherosclerosis is modulated by inflammation and oxidative stress, which play pivotal roles in the pathogenesis of coronary artery disease (CAD), especially in young patients without traditional risk factors for atherosclerosis. In this study, we aimed to study the role of some inflammatory and oxidative stress markers in triggering ST segment elevation myocardial infarction in Egyptian young patients and the potential correlation to the severity of coronary artery lesions in the Egyptian population. Methods: This case-control study recruited 115 premature STEMI patients (aged <45 years) and 55 age-matched healthy controls. Serum CRP, TNF-a, IL-10, and oxidative stress biomarkers [reduced glutathione, total antioxidants, L-ascorbic acid, nitric oxide (NO), and lipid peroxides] were assayed using commercially available kits and correlated with the coronary artery lesion severity, assessed by the SYNTAX score (SS). The ROC curve assessed the biomarker potential of CRP, TNF-a, IL-10, and the TNF-a/IL-10 ratio to discriminate patients from healthy controls. Results: In our study, we found that serum CRP (38.7 ± 1.9 versus 0.8 ± 0.04 mg/L; P < 0.001), TNF-a (68.4 ± 4.7 pg/mL versus 10.8 ± 0.7 pg/mL; P < 0.001), IL-10 (11.9 ± 0.8 pg/mL versus 8.7 ± 0.4 pg/mL; P = 0.001), and TNF-a/IL-10 ratio (6.8 ± 0.5 versus 1.7 ± 0.2; P < 0.001) were significantly higher in patients compared to healthy controls. Regarding oxidative stress markers, serum T-AOC levels (25.5 ± 0.9 versus 14.9 ± 0.4 U/mL; P < 0.001), NO (10.5 ± 0.3 versus 8.1 ± 0.3 nmol/mL; P < 0.001), serum LPO levels (15.8 ± 0.4 versus 4.1 ±0.2 mmol/L; P < 0.001) and lower GSH levels (3.8 ± 0.1 mg/mL versus 5.2 ± 0.2; P < 0.001) were significantly different in patients versus controls. CRP, TNF-a, and NO levels have been significantly correlated with the severity of CAD as assessed by the SYNTAX score. Conclusion: Inflammation and oxidative stress are pathogenically implicated in premature STEMI and correlated with the severity of CAD lesions. The investigated biomarkers can be utilized in risk stratification and are theranostically targetable.