Replication Process Research Articles

HBx promotes tumorigenicity through RRM2-mediated autophagy in hepatocellular carcinoma

BackgroundHepatitis B virus (HBV) infection can exacerbate liver disease progression through multiple mechanisms, eventually leading to hepatocellular carcinoma (HCC). HBV-encoded oncogene X protein (HBx), a key regulatory protein of HBV infection, serves as a positive regulator of hepatocarcinogenesis. The indispensability of the M2 subunit of ribonucleotide-diphosphate reductase (RRM2) lies in its role in facilitating DNA replication and repair processes. In our previous investigation, it was postulated that the gene RRM2 exhibits elevated expression levels in several categories of malignant tumors, particularly in HBV-related HCC. Additionally, it was observed that RRM2 is present within protein complexes that are centered on HBx. In the present investigation, the objective of this work was to investigate the potential relationship between the elevated expression of RRM2 in HBV-related HCC and the influence of HBx on this expression. The study attempted to determine the specific mechanism by which RRM2 is implicated in the promotion of hepatocarcinogenesis by HBx. There have been multiple scholarly proposals suggesting that the induction of autophagy by HBx is a significant intermediary factor in the development of HCC. However, the precise carcinogenic function of HBx-induced autophagy remains a subject of debate.ResultsThis work initially investigated the impact of suppressing cellular autophagy on the malignant biological behaviors of HBx-promoted cells using an in vitro cellular model. The findings revealed that the suppression of cellular autophagy partially disrupted the oncogenic effects of HBx. In light of this, we proceeded to conduct more investigations into the regulatory association between RRM2 and HBx-induced autophagy in the upstream-downstream context. Our data indicate that HBx proteins increase the expression of RRM2. Suppression of RRM2 expression not only hinders HBx-induced autophagy, but also worsens the cellular G1/S blockage and reduces the HBx-induced malignant growth of hepatocellular carcinoma tumors, while stimulating apoptosis.ConclusionsTherefore, we hypothesised that RRM2 is a potential downstream target of HBx-induced hepatocarcinogenesis, and mining the oncogenic mechanism of RRM2 is significant in exploring the preventive treatment of HBV-related HCC.

Identification and expression profiles of granzyme genes in immune response from Nile tilapia (Oreochromis niloticus)

Granzymes (Gzms) are crucial cytotoxic molecules involved in cell-mediated immunity in mammals. However, their roles in teleost fish, such as Nile tilapia, were less understood. In this study, we focused on the identification of granzyme genes (OnGzms) from Nile tilapia (Oreochromis niloticus) and the potential functions in anti-infectious immunity, as well as their expression patterns in response to interleukin-2 stimulation. Eleven Gzm genes were identified in tilapia, including GzmA-1, GzmA-2, GzmK, GzmB-1, GzmB-2, GzmB-3, GzmH-1, GzmH-2, GzmG, GzmF, and GzmE, which contain conserved serine protease active sites. OnGzms were categorized into four groups through homology analysis at a shared locus. Gene replication processes, such as whole-genome replication specific to teleost and unequal crossing over of tandem genes, may lead to multigene copies of OnGzm genes. It appeared that the vertebrate GzmA/K originated from a common ancestor in terms of evolution. After Streptococcus agalactiae, Aeromonas hydrophila, Edwardsiella piscicida and Poly (I:C) challenges, expression of OnGzms was significantly up-regulated only in response to E. piscicida (an intracellular bacterium) infection, indicating potential functions of OnGzms involving in combating intracellular infection by the bacterium E. piscicida. Stimulation with rOnIL-2 led to increased expression of OnGzms in leukocytes, suggesting involvement in cell-mediated cytotoxicity. These findings enriched the knowledge of potential roles in resisting E. piscicida intracellular infection of granzyme genes in teleost fish.

An Experimental Dynamic Investigation of the Influence of Melatonin, Serotonin and Tryptophan on the Stability of the DNA Structure

Background: Small molecules play a crucial role in the exploration of physiological pathways and in drug development by targeting deoxyribonucleic acid (DNA). DNA is a central focus for both endogenous and exogenous ligands, which interact directly or indirectly to regulate transcription and replication processes, thus controlling genetic expression in specific cells. Among these molecules, indole derivatives like tryptophan, serotonin, and melatonin are notable for their widespread presence in nature and significant biological effects. Tryptophan, an essential amino acid, serves as a vital structural element in proteins and a precursor for bioactive compounds like serotonin and melatonin, which impact various physiological functions. Methods: Experimental studies have been conducted to reveal the interaction mechanisms of these endogenous indole derivatives with calf thymus DNA (ct-DNA). These investigations involve viscosity measurements and analysis of double-stranded DNA behavior in the presence of indole molecules, using spectrophotometric UV absorption techniques to assess their impact on DNA stability. Additionally, the influence of calcium and magnesium ions on the resulting complexes of these indole derivatives with ct-DNA has been evaluated. Molecular docking validated our findings, offering additional insights into potential DNA–ligand interactions. Utilizing a crystallographic oligomer with an intercalation gap improved docking accuracy, distinguishing intercalation from groove recognition and enhancing assessment precision. Results: Our study offers detailed insights into the interaction patterns of the indole derivatives with DNA and is highly supported by molecular docking analyses: the indole derivatives were predominantly localized between C and G, interacting via π-π interactions and hydrogen bonds and aligning with known data on conventional intercalators. These findings underscore the importance of small compounds’ planar structure and appropriate size, facilitating tight insertion between adjacent base pairs and disrupting regular DNA stacking. Conclusions: Indoles’ physiological roles and potential as drug candidates targeting specific pathways are highlighted, emphasizing their significance as ubiquitous molecules with the ability to modulate biological effects on DNA structure.

Online Fast Assessment of Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) Proliferation Through Replication

Purpose. A virus, when it infects a host organism, multiplies its ribonucleic acid (RNA) to make it more harmful. We analyze if a view-based temporal template method, the motion history image (MHI) algorithm, may be of use to extract information on the progression of viral interactions over time. Methods. We employ an alternative online processing modality to comprehend and analyze the replication dynamics of severe acute respiratory syndrome coronavirus 2 (SARS CoV 2). The data are evaluated using the computation of a quantitative algorithm, the absolute value of differences (AVD), followed by the generation of temporal template map, the MHI of viral contact. Results. We show that the MHI identifies various stages to examine cultural characteristics and information on the progression of viral interactions over time. The results are more easily obtained compared to a visual inspection of the frames constituting the recordings. MHI also has the advantage of relying on a simple algorithm. The AVD index further provides deeper insights into the dynamics of the viral interaction over time, and its measurement suggests that SARS CoV-2 virus is becoming more widespread. This study shows that the replication process damages the host cell over time, in addition to causing SARS CoV-2 to proliferate. Conclusions. The MHI provides a framework for automated feature extraction from time series data. The combined approach results in decreased computational complexity and enables online assessment of the assay. Clinical and research studies can be conducted to evaluate MHI and AVD and their utility in aiding diagnostic analysis.

RAD52 resolves transcription-replication conflicts to mitigate R-loop induced genome instability

Collisions of the transcription and replication machineries on the same DNA strand can pose a significant threat to genomic stability. These collisions occur in part due to the formation of RNA-DNA hybrids termed R-loops, in which a newly transcribed RNA molecule hybridizes with the DNA template strand. This study investigated the role of RAD52, a known DNA repair factor, in preventing collisions by directing R-loop formation and resolution. We show that RAD52 deficiency increases R-loop accumulation, exacerbating collisions and resulting in elevated DNA damage. Furthermore, RAD52’s ability to interact with the transcription machinery, coupled with its capacity to facilitate R-loop dissolution, highlights its role in preventing collisions. Lastly, we provide evidence of an increased mutational burden from double-strand breaks at conserved R-loop sites in human tumor samples, which is increased in tumors with low RAD52 expression. In summary, this study underscores the importance of RAD52 in orchestrating the balance between replication and transcription processes to prevent collisions and maintain genome stability.

Diverse Head-to-Tail Sequences in the Circular Genome of Human Bocavirus Genotype 1 among Children with Acute Respiratory Infections Implied the Switch of Template Chain in the Rolling-Circle Replication Model.

Head-to-tail sequences have been reported in human bocavirus (HBoV) 1-4. To reveal their features and functions, HBoV DNA was screened among respiratory specimens from pediatric patients with an acute respiratory infection (ARI) between April 2020 and December 2022, followed by HBoV genotyping. Head-to-tail sequences were detected using nested PCR, TA cloning, and Sanger sequencing, and these findings were confirmed by mNGS and amplicon sequencing. The secondary structure was predicted using the Mfold web server. The results indicated that head-to-tail sequences were detected in 42 specimens through TA cloning from 351 specimens positive for HBoV1 DNA, yielding 92 sequences into 32 types and 2 categories. Additionally, head-to-tail sequences were detected in 16 specimens by amplicon sequencing, yielding 60 sequences categorized into 23 types. The 374nt type, detected in 13 specimens, contains variants 374a and 374b, which differ in the unpaired loop regions of the palindrome or complementary reverse sequences, implying a switch of template chains during the replication process. The mNGS results in three specimens confirmed the presence of circular genome in copies below 1%. In conclusion, head-to-tail sequences of HBoV1 were common in children with ARI and were highly diverse in length and sequences. The variants may be generated by the switch of the template chain in the rolling-circle replication model.

TDFFM: Transformer and Deep Forest Fusion Model for Predicting Coronavirus 3C-Like Protease Cleavage Sites.

COVID-19, caused by the highly contagious SARS-CoV-2 virus, is distinguished by its positive-sense, single-stranded RNA genome. A thorough understanding of SARS-CoV-2 pathogenesis is crucial for halting its proliferation. Notably, the 3C-like protease of the coronavirus (denoted as 3CLpro) is instrumental in the viral replication process. Precise delineation of 3CLpro cleavage sites is imperative for elucidating the transmission dynamics of SARS-CoV-2. While machine learning tools have been deployed to identify potential 3CLpro cleavage sites, these existing methods often fall short in terms of accuracy. To improve the performances of these predictions, we propose a novel analytical framework, the Transformer and Deep Forest Fusion Model (TDFFM). Within TDFFM, we utilize the AAindex and the BLOSUM62 matrix to encode protein sequences. These encoded features are subsequently input into two distinct components: a Deep Forest, which is an effective decision tree ensemble methodology, and a Transformer equipped with a Multi-Level Attention Model (TMLAM). The integration of the attention mechanism allows our model to more accurately identify positive samples, thus enhancing the overall predictive performance. Evaluation on a test set demonstrates that our TDFFM achieves an accuracy of 0.955, an AUC of 0.980, and an F1-score of 0.367, substantiating the model's superior prediction capabilities.

Wildland-Urban Interface fire exposure of rural settlements: The case of Montesinho Natural Park

Coexistence with wildfires has always been part of human life. However, the severity of these events has intensified due to the combined effects of climate change and interventions on the environment. Rural settlements, especially those with heritage values, are vulnerable, leaving their existence under constant threat. Montesinho Natural Park, characterised by many settlements with remarkable vernacular buildings, exemplifies this situation. In response to this challenge and added to the wildfire hazard, it was chosen to apply the Wildland-Urban Interface Index (WUIX) methodology to assess the exposure of the wildland-urban interface (WUI) to wildfires in three of the villages: Aveleda, Montesinho, and Rio de Onor. The WUI represents the boundary between the natural landscape and the community and might serve as a path for the percolation of fire. The study focused on evaluating the evolution of WUI over time for those three villages by taking advantage of the two open data national orthophotos of 2018 and 2021. The initial stage of vegetation recognition was changed to an automated process, relying on a Python script that captures the image for the database, extracts the vegetation, converts, and prepares the files for applying WUIX, and again converts and georeferenced the results for analysis. This approach makes the application a faster, reliable and replicable process. As a result, maps of WUIX affectation were generated, visually and quantitatively pointing to the village of Aveleda as the one with a higher level of exposure. While the results alone cannot support the entire vulnerability assessment of those villages, they serve as a complement to efforts to define proactive measures for safeguarding the rural heritage.

Analyzing Telomeric Protein-DNA Interactions Using Single-Molecule Magnetic Tweezers.

Telomeres, the protective structures at the ends of chromosomes, are crucial for maintaining cellular longevity and genome stability. Their proper function depends on tightly regulated processes of replication, elongation, and damage response. The shelterin complex, especially Telomere Repeat-binding Factor 1 (TRF1) and TRF2, plays a pivotal role in telomere protection and has emerged as a potential anti-cancer target for drug discovery. These proteins bind to the repetitive telomeric DNA motif TTAGGG, facilitating the formation of protective structures and recruitment of other telomeric proteins. Structural methods and advanced imaging techniques have provided insights into telomeric protein-DNA interactions, but probing the dynamic processes requires single-molecule approaches. Tools like magnetic tweezers, optical tweezers, and atomic force microscopy (AFM) have been employed to study telomeric protein-DNA interactions, revealing important details such as TRF2-dependent DNA distortion and telomerase catalysis. However, the preparation of single-molecule constructs with telomeric repetitive motifs continues to be a challenging task, potentially limiting the breadth of studies utilizing single-molecule mechanical methods. To address this, we developed a method to study interactions using full-length human telomeric DNA with magnetic tweezers. This protocol describes how to express and purify TRF2, prepare telomeric DNA, set up single-molecule mechanical assays, and analyze data. This detailed guide will benefit researchers in telomere biology and telomere-targeted drug discovery.

SARS-CoV-2 JN.1 variant: a short review.

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a single-stranded, positive-sense RNA virus. The SARS-CoV-2 virus is evolving continuously, and many variants have been detected over the last few years. SARS-CoV-2, as an RNA virus, is more prone to mutating. The continuous evolution of the SARS-CoV-2 virus is due to genetic mutation and recombination during the genomic replication process. Recombination is a naturally occurring phenomenon in which two distinct viral lineages simultaneously infect the same cellular entity in an individual. The evolution rate depends on the rate of mutation. The rate of mutation is variable among the RNA viruses, with the SARS-CoV-2 virus exhibiting a lower rate of mutation than other RNA viruses. The novel 3'-to-5' exoribonuclease proofreading machinery is responsible for a lower rate of mutation. Infection due to the SARS-CoV-2, influenza, and respiratory syncytial virus has been reported from around the world during the same period of fall and winter, resulting in a "tripledemic." The JN.1 variant, which evolved from the predecessor, the omicron variant BA.2.86, is currently the most dominant globally. The impact of the JN.1 variant on transmissibility, disease severity, immune evasion, and diagnostic and therapeutic escape will be discussed.

Developing a range-wide sampling framework for endangered species: a case study with light-footed Ridgway’s rail

Abstract Monitoring provides the foundation for evaluating recovery of endangered species, yet many species lack monitoring programs designed to integrate a species’ unique attributes, specific monitoring objectives, and principles of statistical sampling theory. We developed a framework for monitoring and assessment of endangered light-footed Ridgway’s rails (Rallus obsoletus levipes) across their U.S. range, relative to multi-scale recovery goals. We created spatially explicit sample units and a sampling frame covering all potential habitat to facilitate range-wide probability sampling, and also built a model of the call-broadcast process commonly used to survey marsh birds that included heterogeneity in availability for detection and conditional detectability for each bird during each survey. We used the model to simulate 96 sampling strategies that included different levels of replication, multiple approaches for sample allocation amongst strata, and both simple random and weighted probability sampling (i.e., weights proportional to local rail abundance) of sample units within strata. Effective monitoring surveyed ≥ 20–30% of the sampling frame on ≥ 3 occasions, with weighted sample selection and more targeted sampling (50% of units) for strata that are key to species recovery. We also tested Bayesian N-mixture models for estimating abundance and show that multiple models provide reasonable estimates. This work lays the foundation for statistical sampling and multi-scale population estimation for an endangered bird, and for refinement of abundance estimation models. Moreover, this work provides a replicable process for building customized and statistically defensible sampling frameworks to assess recovery of endangered species that can be used for other sensitive species.

Hepatitis B Viral Protein HBx: Roles in Viral Replication and Hepatocarcinogenesis.

Hepatitis B virus (HBV) infection remains a major public health concern worldwide, with approximately 296 million individuals chronically infected. The HBV-encoded X protein (HBx) is a regulatory protein of 17 kDa, reportedly responsible for a broad range of functions, including viral replication and oncogenic processes. In this review, we summarize the state of knowledge on the mechanisms underlying HBx functions in viral replication, the antiviral effect of therapeutics directed against HBx, and the role of HBx in liver cancer development (including a hypothetical model of hepatocarcinogenesis). We conclude by highlighting major unanswered questions in the field and the implications of their answers.

Computational investigation of novel synthetic analogs of C-1′β substituted remdesivir against RNA-dependent RNA-polymerase of SARS-CoV-2

Remdesivir, a C-nucleotide prodrug binds to the viral RNA-dependent-RNA polymerase (RdRp) and inhibits the viral replication by terminating RNA transcription prematurely. It is reported in literature that interaction between the C-1’β–CN moiety of Remdesivir (RDV) and the Ser861 residue in RdRp enzyme, causes a delayed chain termination during the RNA replication process and is one of the important aspect of its mechanism of action. In the pursuance of increasing the biological activity of RDV and enhancing the SAR studies, against RNA viruses, we have designed its fourteen C1’β substituted analogs, 10 –23 bearing 4/5-membered heterocyclic rings. The docking and 100 ns molecular dynamics (MD) simulations of 10-23 to the RdRp protein (PDB ID: 7L1F) revealed important interactions between 2’,3’-diol, oxo group of phosphoramidate, nitrogen residues of heterocyclic rings of synthetic molecules with Arg555, Arg553, Ser759, Cys622, Asn691, Asp623 amino acid residues of protein. The docking score of 2-ethylbutyl ((S)-(((2R,3S,4R,5R)-5-(4-aminopyrrolo[2,1-f][1,2,4]triazin-7-yl)-3,4-dihydroxy-5-(1H-1,2,3-triazol-4-yl)tetrahydrofuran-2-yl)methoxy)(phenoxy)phosphoryl)-L-alaninate, 11 was found to be the higher than RDV among 14 new compounds i.e. -5.20 kcal/mol. Out of 3 compounds, 10, 12 and 13 submitted for MD simulations and Molecular Mechanics Poisson-Boltzmann Surface Area (MMPBSA) analysis, trifluoro-oxadiazole derivative, 13 showed higher binding energy as compared to Remdesivir. The predicted ADMET properties of 14 compounds showed their potential for being drug candidates. The present study suggests that substitution at the C1’β position by 4/5-membered rings plays an important role in the interactions between nucleoside/tide and target protein.

Etravirine Prevents West Nile Virus and Chikungunya Virus Infection Both In Vitro and In Vivo by Inhibiting Viral Replication.

Diseases transmitted by arthropod-borne viruses such as West Nile virus (WNV) and chikungunya virus (CHIKV) pose threat to global public health. Unfortunately, to date, there is no available approved drug for severe symptoms caused by both viruses. It has been reported that reverse transcriptase inhibitors can effectively inhibit RNA polymerase activity of RNA viruses. We screened the anti-WNV activity of the FDA-approved reverse transcriptase inhibitor library and found that 4 out of 27 compounds showed significant antiviral activity. Among the candidates, etravirine markedly inhibited WNV infection in both Huh 7 and SH-SY5Y cells. Further assays revealed that etravirine inhibited the infection of multiple arboviruses, including yellow fever virus (YFV), tick-borne encephalitis virus (TBEV), and CHIKV. A deeper study at the phase of action showed that the drug works primarily during the viral replication process. This was supported by the strong interaction potential between etravirine and the RNA-dependent RNA polymerase (RdRp) of WNV and alphaviruses, as evaluated using molecular docking. In vivo, etravirine significantly rescued mice from WNV infection-induced weight loss, severe neurological symptoms, and death, as well as reduced the viral load and inflammatory cytokines in target tissues. Etravirine showed antiviral effects in both arthrophlogosis and lethal mouse models of CHIKV infection. This study revealed that etravirine is an effective anti-WNV and CHIKV arbovirus agent both in vitro and in vivo due to the inhibition of viral replication, providing promising candidates for clinical application.

Accountant Skill Gen Z Fresh Graduates in Digital Era: A Systematic Literature Review

Purpose: The purpose of this study is to investigate the impact of emerging technologies on the skills required for accountants in the digital era, particularly focusing on the skills needed by Gen Z fresh graduates. Method: The present study builds on a systematic literature review of 41 academic articles published in Scopus Database between 2017 and 2024. It is based on the “preferred reporting items for systematic reviews and meta-analyses” (PRISMA) method. Synthesising research through a transparent, rigorous and replicable process makes it possible to explore and identify accountant skill in digital era. Findings: The findings highlighted the need for accountants to possess digital literacy skills, data analytics skills, and the ability to work collaboratively with professionals from other fields. The study also emphasized the importance of continuous professional development to keep pace with the rapid technological advancements in accounting. Originality/Value: This paper offers a comprehensive literature review on impact of emerging technologies on the skills required for accountants in the digital era, particularly focusing on the skills needed by Gen Z fresh graduates. It could provide a insight and guidance for accounting fresh graduate to possess digital literacy skills, data analytics skills, and soft skills to adapt to technological changes and meet the needs of the dynamic accounting industry.

A INTERAÇÃO ENTRE COVID-19 E O SISTEMA IMUNOLÓGICO: MECANISMOS E IMPLICAÇÕES CLÍNICAS

The infection by SARS-CoV-2, the virus responsible for COVID-19, triggers a series of immune responses in the host organism. These responses, while essential for pathogen elimination, can also cause significant damage to host tissues. This work details the mechanism of SARS-CoV-2 infection, from its structure to the processes of viral entry and replication, as well as the mechanisms related to infection and activation of the immune system, which can culminate in a cytokine storm. Understanding these mechanisms is crucial for developing effective therapeutic strategies, vaccines, and alternative therapies that were urgently released during the pandemic. Continuous research, especially related to the immune response, is necessary to enhance knowledge and preparedness for future pandemics.

Structures of the mitochondrial single-stranded DNA binding protein with DNA and DNA polymerase γ.

The mitochondrial single-stranded DNA (ssDNA)binding protein, mtSSB or SSBP1, binds to ssDNA to prevent secondary structures of DNA that could impede downstream replication or repair processes. Clinical mutations in the SSBP1 gene have been linked to a range of mitochondrial disorders affecting nearly all organs and systems. Yet, the molecular determinants governing the interaction between mtSSB and ssDNA have remained elusive. Similarly, the structural interaction between mtSSB and other replisome components, such as the mitochondrial DNA polymerase, Polγ, has been minimally explored. Here, we determined a 1.9-Å X-ray crystallography structure of the human mtSSB bound to ssDNA. This structure uncovered two distinct DNA binding sites, a low-affinity site and a high-affinity site, confirmed through site-directed mutagenesis. The high-affinity binding site encompasses a clinically relevant residue, R38, and a highly conserved DNA base stacking residue, W84. Employing cryo-electron microscopy, we confirmed the tetrameric assembly in solution and capture its interaction with Polγ. Finally, we derived a model depicting modes of ssDNA wrapping around mtSSB and a region within Polγ that mtSSB binds.

TDSQL: Tencent Distributed Database System

Distributed databases have become indispensable in contemporary computing and data processing, owing to their pivotal role in ensuring high availability and scalability. They effectively cater to the requirements of data management and high-concurrency access. However, developing a distributed database system that is well-suited for diverse application scenarios, particularly for large-scale applications, presents several challenges. These challenges include ensuring data consistency and achieving high levels of performance. This paper presents TDSQL, a distributed database system that prioritizes core design principles of distributed systems, including high availability, strong consistency, and scalability. In particular, TDSQL has achieved high performance through over a decade of practical experience and optimization in various modules, such as the kernel, synchronous replication, and transaction processing, in large-scale application scenarios. By conducting the TPC-C benchmark test, TDSQL demonstrated outstanding performance, achieving a throughput of 814 million tpmC across 1650 database nodes, with a jitter rate of less than 0.2%. This jitter rate is an order of magnitude lower than the standard required, showcasing the system's stability and reliability. During the 8-hour TPC-C standard stress test, TDSQL successfully completed over 860 billion transactions and processed 40 trillion order details, with zero forced rollbacks and zero data inconsistency.

Unlocking Secrets: Lipid Metabolism and Lipid Droplet Crucial Roles in SARS-CoV-2 Infection and the Immune Response.

Lipid droplets (LD) are crucial for maintaining lipid and energy homeostasis within cells. LDs are highly dynamic organelles that present a phospholipid monolayer rich in neutral lipids. Additionally, LDs are associated with structural and non-structural proteins, rapidly mobilizing lipids for various biological processes. Lipids play a pivotal role during viral infection, participating during viral membrane fusion, viral replication, and assembly, endocytosis, and exocytosis. Severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) infection often induces LD accumulation, which is used as a source of energy for the replicative process. These findings suggest that LDs are a hallmark of viral infection, including SARS-CoV-2 infection. Moreover, LD participates in the inflammatory process and cell signaling, activating pathways related to innate immunity and cell death. Accumulating evidence demonstrates that LD induction by SARS-CoV-2 is a highly coordinated process, aiding replication and evading the immune system, and may contribute to the different cell death process observed in various studies. Nevertheless, recent research in the field of LDs suggests these organelles according to the pathogen and infection conditions may also play roles in immune and inflammatory responses, protecting the host against viral infection. Understanding how SARS-CoV-2 influences LD biogenesis is crucial for developing novel drugs or repurposing existing ones. By targeting host lipid metabolic pathways exploited by the virus, it is possible to impact viral replication and inflammatory responses. This review seeks to discuss and analyze the role of LDs during SARS-CoV-2 infection, specifically emphasizing their involvement in viral replication and the inflammatory response.

1mΨ influences the performance of various positive-stranded RNA virus-based replicons

Self-amplifying RNAs (saRNAs) are versatile vaccine platforms that take advantage of a viral RNA-dependent RNA polymerase (RdRp) to amplify the messenger RNA (mRNA) of an antigen of interest encoded within the backbone of the viral genome once inside the target cell. In recent years, more saRNA vaccines have been clinically tested with the hope of reducing the vaccination dose compared to the conventional mRNA approach. The use of N1-methyl-pseudouridine (1mΨ), which enhances RNA stability and reduces the innate immune response triggered by RNAs, is among the improvements included in the current mRNA vaccines. In the present study, we evaluated the effects of this modified nucleoside on various saRNA platforms based on different viruses. The results showed that different stages of the replication process were affected depending on the backbone virus. For TNCL, an insect virus of the Alphanodavirus genus, replication was impaired by poor recognition of viral RNA by RdRp. In contrast, the translation step was severely abrogated in coxsackievirus B3 (CVB3), a member of the Picornaviridae family. Finally, the effects of 1mΨ on Semliki forest virus (SFV), were not detrimental in in vitro studies, but no advantages were observed when immunogenicity was tested in vivo.