Talat Mokhtari-Azad

Context: Ferroptosis is an understudied iron-dependent cell death pathway characterized by lipid peroxidation and has emerged as a critical mediator of virus-induced cytotoxicity and inflammation, particularly in respiratory viral infections. Objectives: This study examined the temporal regulation of ACSL4, a key pro-ferroptotic enzyme. Evidence Acquisition: A549 cells were infected with influenza A/H1N1, influenza A/H3N2, and influenza B viruses and assessed at 24 and 48 hours post-infection (hpi). ACSL4 expression was evaluated using quantitative reverse transcription-polymerase chain reaction, followed by post hoc analysis. Results: ACSL4 expression showed modest changes at 48 hpi in the A/H1N1 and A/H3N2 groups. However, a pronounced upregulation was observed at 24 hpi, particularly in the A/H1N1 group, with a log fold change increase of 1.52 (P = 0.005). The greatest downregulation was observed at 48 hpi in the influenza B group, with a significant log fold change of -4.11. Conclusions: These findings indicate the time-dependent activation of pro-ferroptotic mRNA associated with ferroptosis during influenza virus infection and suggest that ACSL4 may be a mediator of virus-induced cell death and a target for antiviral interventions.

Background: Influenza viruses induce multiple programmed cell death pathways in respiratory epithelial cells. These pathways are associated with host antiviral defense but may also contribute to tissue damage through inflammation and increased disease severity, thereby functioning as a double-edged sword. Apoptosis is a major cell death pathway that regulates the fate of infected cells following viral infection. Objectives: This study evaluated the mRNA expression of BAX, BAK1, and Bcl-2, three key apoptosis-regulating genes, in A549 lung epithelial cells infected with influenza A/H1N1, A/H3N2, and influenza B viruses. Methods: A549 cells were cultured and inoculated with 100 TCID50 of influenza A/H1N1, A/H3N2, and influenza B viruses for 48 hours. Results were compared with those for uninfected control cells. At 48 hours post-infection, RNA extraction (RNJIA, ROJE) and cDNA synthesis were performed, and the mRNA expression levels of BAX, BAK1, and Bcl-2 were assessed using SYBR Green reverse transcription quantitative PCR. All analyses were based on log fold change (logFC). Differential gene expression versus control was evaluated using the Wilcoxon signed-rank test and a one-sample t test, whereas time-dependent changes within each virus group were assessed using the Kruskal-Wallis test and one-way analysis of variance, followed by pairwise post-hoc tests with Benjamini-Hochberg correction. Results: At 48 hours post-infection, BAX expression was significantly upregulated in A/H1N1-infected cells (log fold change = 2.85, P = 0.031). Although A/H3N2 also upregulated BAX expression (log fold change = 2.35, P = 0.061), this change was not significant. Minor downregulation of BAX was observed in influenza B-infected cells (log fold change = -0.26, P = 0.463). BAK1 expression was significantly upregulated in A/H3N2-infected cells (log fold change = 6.20, P = 0.019). A/H1N1-infected cells also showed BAK1 upregulation (log fold change = 2.10, P = 0.431), although this change was not significant, whereas influenza B-infected cells showed significant BAK1 upregulation (log fold change = 6.21, P = 0.025). Bcl-2 was significantly downregulated in all three infected groups (A/H1N1: log fold change = -6.05, P = 0.040; A/H3N2: log fold change = -4.02, P = 0.002; influenza B: log fold change = -0.70, P = 0.032). Conclusions: This experiment demonstrated strain-dependent regulation of apoptosis-related genes in A549 cells following infection with different influenza viruses. Significant upregulation of BAX and BAK1 after A/H1N1 infection, upregulation of BAK1 after A/H3N2 and influenza B infection, and downregulation of Bcl-2 in all infected groups suggest activation of pro-apoptotic signaling. These findings are preliminary transcriptional observations from an in vitro system and may provide insight into mechanisms involved in the regulation of pro-apoptotic genes in the apoptosis pathway.

Introduction: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been a significant public health issue worldwide in recent years. The most recently circulating variant of SARS-CoV-2, Omicron, and its rapid evolution into various sub-lineages have raised concerns regarding the effects of the immunity on the virus epitopes, in the human population. The present study evaluated and compared these important variations among different Omicron sub-lineages in Iran.Methodology: From October 2023 to August 2024, high coverage whole genome sequences of 49 SARS-CoV-2 strains were subjected to phylogenetic analysis and evaluation of B cell, CD4⁺, and CD8⁺ T cell epitopes in Iran National Influenza Centre.Results: The phylogenetic tree exhibited eight Nextstrain clades (21L, 22F, 23B, 23H, 23D, 24A, 24B, and 24C) in 48 studied strains, and one recombinant strain (XDK.1). The evaluation of B cell, CD4⁺, and CD8⁺ T cell epitopes in all studied strains revealed 31, 65, and 78%, of conservation, respectively. The low B cell epitopes conservation rate among Omicron sub-lineages underscored the escaping from neutralizing humoral immunity. T cell epitopes of the SARS-CoV-2 were considerably preserved across major Omicron sub-lineages. Conservation levels varied based on the epitope class (higher for CD8⁺ vs. CD4⁺), protein (higher for non-spike vs. spike), and clades (higher for 21L, 22F, 23B, 23H, 23D, and 24B vs. 24A and 24C).Conclusion: Herein, the increased conservation of CD8⁺ epitopes compared to CD4⁺ and B cell epitopes is probably attributable to the shorter length of the peptides associated with CD8⁺ epitopes. The high rate of T-cell epitopes conservation in non-spike proteins among different sub-lineages of the Omicron in this study highlighted the importance of cell-mediated immunity and suggested that non-spike proteins might be more attractive targets for future SARS-CoV-2 vaccines.

Measles, an ongoing public health concern, demands continuous molecular surveillance and virus characterization for elimination. Despite Iran achieving measles elimination status in 2019 through robust molecular testing and vaccination, the COVID-19 pandemic disrupted global vaccination efforts, leading to increased measles-related morbidity and mortality. This study aims to overview measles virus serological and molecular traits in Iran from 1st January 2021 to 30th April 2023. Following World Health Organization and Center for Diseases and Control protocols, serological tests were performed on suspected cases and the nucleoprotein (N) gene of confirmed cases were subsequently amplified using molecular methods and were sequenced afterwards for measles genotyping. Phylogenetic analysis was performed with the obtained sequences. Analyzing 17,343 suspected cases from 1st January 2021 to 30th April 2023, 936, 177, and 164 samples were positive using ELISA, quantitative Reverse transcription PCR, and Reverse transcription PCR, respectively. The B3 genotype predominated, notably in Iran’s South East (41%), Central (28%), and South (13%) regions. Provinces bordering countries with measles outbreaks exhibited higher risk of virus importation. Genetic comparisons with measles sequences submitted to NCBI and MeaNS databases revealed direct importation and contact transmission. Regular surveillance and genetic analysis are critical for understanding measles transmission and reacting to outbreaks. The COVID-19 pandemic yielded mixed effects on measles cases, enhancing hygiene measures while causing underreporting and vaccination gaps. Vigilance against measles resurgence is crucial, requiring cross-border transmission studies, improving cross-border surveillance and adaptable vaccination strategies.

Key clinical message: In SARS‐CoV‐2 pandemic different disorders in coagulation pathways in COVID‐19 patients were reported. We described a 44‐year‐old female with COVID‐19 and protein C deficiency history. She did not show any coagulation disorder during her disease course. Complete genome sequencing of SARS‐CoV‐2 was performed and some mutations identified and compared with Wuhan strain. Besides hospitalized patients, in COVID‐19 outpatients with low concentration of protein C, early prescription of an anticoagulant such as heparin could be helpful in prevention of venous thromboembolism or pulmonary embolism.

Background: Human orthopneumovirus (HOPV) or respiratory syncytial virus (RSV) is one of the important causes of acute respiratory infections (ARIs) during the cold months of the year worldwide. Many countries have reported an absence of ARIs due to HOPV during the winter of 2020–2021 associated with preventive measures to reduce the spread of SARS-CoV2. However, with the reduction of COVID-19 public health restrictions and the absence of immunity in the community due to the lack of exposure in the previous season, many countries had a delayed HOPV outbreak. Here we reported the impact of COVID-19 on the changing pattern of HOPV infection in Iran. Methods: Throat and nasopharyngeal swab samples were collected from patients (children and adults) with ARIs and sent to the Iran National Influenza Center. After RNA extraction, Real time RT-PCR was performed for HOPV detection. Results: In 260 samples collected from patients with ARIs in three different groups, which included children in March 2021, pilgrims in July 2022, and outpatients during November and December 2022, no HOPV was detected in any group. Conclusions: The lack of HOPV activity in Iran during the winter of 2020–2021 and then the resurgence in spring 2022 and again the absence of activity in summer and autumn 2022 was extraordinary in the HOPV epidemiology, and probably due to the implementation of public health non-pharmaceutical interventions to reduce the spread of SARS-CoV2. Although it is not possible to keep such restrictions, similar methods can be taken to control outbreaks caused by respiratory viruses.

Background and Aims: Real‐time reverse‐transcriptase polymerase chain reaction (real‐time RT‐PCR) is the gold standard test for diagnosis of severe acute respiratory syndrome coronavirus 2 (SARS‐CoV‐2). However, when the test result is near the detection limit of the assay the possibility of getting false positive or negative results is high. In addition, it might result in single target gene positive (STGP) results which should be interpreted with caution.Methods: This study was performed on 29,962 nasal swabs from July 1 to August 31, 2020. Ct values less than 40 for each or both of N and RdRp genes were recommended to be selected as positive. Positive samples for one gene with the Cts more than 35 were rechecked by adding more templates.Results: The results showed that 1016 (3.39%) samples were positive just for one gene with high Ct values. The results of the second reactions showed that 325 (31.99%) samples were positive for both N and RdRp which were reported positive, 301 (29.65%) were positive only for one gene which were considered as suspicious cases and resampling was suggested for them. Finally, 390 (38.385%) samples were negative for both genes.Conclusion: In conclusion, tracking weak positive results of SARS‐CoV‐2 real‐time RT‐PCR revealed that most of the individuals who were STGP clean the infection completely in less than a week which showed they were in the convalescent phase of infection. However, some of them who were in the beginning of infection showed a decrease in Ct value during a week, so they could spread the virus in the society.