Viral DNA Research Articles

Global Re-emergence of Monkeypox: A Synoptic Review

AbstractAs the coronavirus disease 2019 pandemic continues to rage, the unprecedented manifold increase in monkeypox (mpox) cases throughout the world is raising qualms about a possible pandemic. As of January 2024, the disease has been reported in around 116 countries, with nearly 92,500 confirmed cases and 170 deaths. In this mini-review, we have endeavored to cover multiple aspects of the mpox disease. Mpox virus is an enveloped double-stranded DNA virus that belongs to the Orthopoxvirus genus. The disease is endemic in certain African countries. Until recently, however, the disease was rarely reported in Europe and the United States. In contrast to the previous outbreaks outside Africa, reported cases in the 2022 outbreak did not travel to the endemic areas. Superspreading events at mass gatherings, multiple-partner sexual encounters, and international travel were major drivers of the recent global outbreak of mpox. The 2022 mpox virus may have undergone accelerated evolution. It diverges from the related 2018 to 2019 viruses by around 50 single-nucleotide polymorphisms, some of which brought about amino acid changes in immunogenic surface glycoprotein B21. Differential diagnosis for mpox could be quite challenging since it can masquerade as a wide variety of illnesses. Worse still, some patients may be asymptomatic or show subtle symptoms. The infection is confirmed by conventional or real-time polymerase chain reaction on lesion material. Although there is no specific therapy approved for mpox infections, two antivirals (tecovirimat and brincidofovir) and vaccinia immune globulin may be used. Vaccines also provide protection against mpox when properly administered prior to exposure. Finally, the implementation of preventive measures is of paramount importance, especially in regions where mpox transmission is widespread and among high-risk populations.

A critical review of diagnostic and prognostic markers of chronic hepatitis B infection.

A major worldwide health concern, chronic hepatitis B necessitates precise prognostic and diagnostic indicators for clinical guidance. This article highlights the clinical importance and current issues of the major markers used in both the detection and prognosis of chronic hepatitis B. An important indicator of an ongoing and persistent infection is the hepatitis B surface antigen. Hepatitis B virus DNA quantification monitoring aids in assessing viral load and hepatic cancer risk. While limited evidence of liver damage is provided by alanine aminotransferase levels, the hepatitis B core antibody verifies acute infection. Seroconversion to the hepatitis B e antibody is linked to a lower risk of disease development, and the hepatitis B e antigen status is a critical prognostic factor. Treatment choices are guided by a biopsy of the liver or minimally invasive liver fibrosis detection. Genotypes of the hepatitis B virus and host variables influence the prognosis by adding to the disease's variability. Noninvasive techniques to evaluate the severity of the disease are provided by serum markers of fibrosis, such as the fibrosis score based on four criteria and the aspartate aminotransferase-to-platelet ratio index. The requirement for indicators that distinguish between distinct viral phases and increase specificity in evaluating liver damage is one of the challenges facing chronic hepatitis B research. Even though it is quite difficult to find reliable biomarkers for resistance especially when it comes to hepatocellular cancer risk estimation, there are advanced methods, which include imaging and omics that can help in improving the accuracy of the diagnostics and prognosis. Interventions early point that improve patient outcomes are made possible using diagnostics and prognostics as they are quite effective in managing the complicated landscape of chronic hepatitis B. Key in addressing these challenges today and improving the diagnostic and prognostic markers in thefuture, particularly those that would support the development of successful treatment plans for people living with chronic hepatitis B virus (HBV), are scientific research, technological advances and collaborations.

Experimental inoculation of pigs with monkeypox virus results in productive infection and transmission to sentinels

ABSTRACT Monkeypox virus (MPXV) is a re-emerging zoonotic poxvirus responsible for producing skin lesions in humans. Endemic in sub-Saharan Africa, the 2022 outbreak with a clade IIb strain has resulted in ongoing sustained transmission of the virus worldwide. MPXV has a relatively wide host range, with infections reported in rodent and non-human primate species. However, the susceptibility of many domestic livestock species remains unknown. Here, we report on a susceptibility/transmission study in domestic pigs that were experimentally inoculated with a 2022 MPXV clade IIb isolate or served as sentinel contact control animals. Several principal-infected and sentinel contact control pigs developed minor lesions near the lips and nose starting at 12 through 18 days post-challenge (DPC). No virus was isolated and no viral DNA was detected from the lesions; however, MPXV antigen was detected by IHC in tissue from a pustule of a principal infected pig. Viral DNA and infectious virus were detected in nasal and oral swabs up to 14 DPC, with peak titers observed at 7 DPC. Viral DNA was also detected in nasal tissues or skin collected from two principal-infected animals at 7 DPC post-mortem. Furthermore, all principal-infected and sentinel control animals enrolled in the study seroconverted. In conclusion, we provide the first evidence that domestic pigs are susceptible to experimental MPXV infection and can transmit the virus to contact animals.

Characterizing Glycosylation of Adeno-Associated Virus Serotype 9 Capsid Proteins Generated from HEK293 Cells through Glycopeptide Mapping and Released Glycan Analysis.

Recombinant adeno-associated viral (AAV) vectors have emerged as prominent gene delivery vehicles for gene therapy. AAV capsid proteins determine tissue specificity and immunogenicity and play important roles in receptor binding, the escape of the virus from the endosome, and the transport of the viral DNA to the nuclei of target cells. Therefore, the comprehensive characterization of AAV capsid proteins is necessary for a better understanding of the vector assembly, stability, and transduction efficiency of AAV gene therapies. Glycosylation is one of the most common post-translational modifications (PTMs) and may affect the tissue tropism of AAV gene therapy. However, there are few studies on the characterization of the N- and O-glycosylation of AAV capsid proteins. In this study, we identified the N- and O-glycosylation sites and forms of AAV9 capsid proteins generated from HEK293 cells using liquid chromatography-tandem mass spectrometry (LC-MS)-based glycopeptide mapping and identified free N-glycans released from AAV9 capsid proteins by PNGase F using hydrophilic interaction (HILIC) LC-MS and HILIC LC-fluorescence detection (FLD) methods. This study demonstrates that AAV9 capsids are sprinkled with sugars, including N- and O-glycans, albeit at low levels. It may provide valuable information for a better understanding of AAV capsids in supporting AAV-based gene therapy development.

Individualized radiology screening for newly diagnosed nasopharyngeal carcinoma

ObjectivesCurrent guidelines recommend universal PET/CT screening for metastases staging in newly diagnosed nasopharyngeal carcinoma (NPC) despite the low rate of synchronous distant metastasis (SDM). The study aims to achieve individualized screening recommendations of NPC based on the risk of SDM. Methods and materials18 pre-treatment peripheral blood indicators was retrospectively collected from 2271 primary NPC patients. A peripheral blood risk score (PBRS) was constructed by indicators associated with SDM on least absolute shrinkage and selection operator (LASSO) regression. The PBRS-based distant metastases (PBDM) model was developed from features selected by logistic regression analyses in the training cohort and then validated in the validation cohort. Receiver operator characteristic curve analysis, calibration curves, and decision curve analysis were applied to evaluate PBDM model performance. ResultsPre-treatment Epstein–Barr viral DNA copy number, percentage of total lymphocytes, serum lactate dehydrogenase level, and monocyte-to-lymphocyte ratio were most strongly associated with SDM in NPC and used to construct the PBRS. Sex (male), T stage (T3-4), N stage (N2-3), and PBRS (≥1.076) were identified as independent risk factors for SDM and applied in the PBDM model, which showed good performance. Through the model, patients in the training cohort were stratified into low-, medium-, and high-risk groups. Individualized screening recommendations were then developed for patients with differing risk levels. ConclusionThe PBDM model offers individualized recommendations for applying PET/CT for metastases staging in NPC, allowing more targeted screening of patients with greater risk of SDM compared with current recommendations.

Partial Alleviation of Homologous Superinfection Exclusion of SeMNPV Latently Infected Cells by G1 Phase Infection and G2/M Phase Arrest.

Viral infection can regulate the cell cycle, thereby promoting viral replication. Hijacking and altering the cell cycle are important for the virus to establish and maintain a latent infection. Previously, Spodoptera exigua multiple nucleopolyhedrovirus (SeMNPV)-latently infected P8-Se301-C1 cells, which grew more slowly than Se301 cells and interfered with homologous SeMNNPV superinfection, were established. However, the effects of latent and superinfection with baculoviruses on cell cycle progression remain unknown. In this study, the cell cycle profiles of P8-Se301-C1 cells and SeMNPV or Autographa californica multiple nucleopolyhedrovirus (AcMNPV)-infected P8-Se301-C1 cells were characterized by flow cytometry. The results showed that replication-related genes MCM4, PCNA, and BAF were down-regulated (p < 0.05) in P8-Se301-C1 cells, and the S phase of P8-Se301-C1 cells was longer than that of Se301 cells. P8-Se301-C1 cells infected with SeMNPV did not arrest in the G2/M phase or affect the expression of Cyclin B and cyclin-dependent kinase 1 (CDK1). Furthermore, when P8-Se301-C1 cells were infected with SeMNPV after synchronized treatment with hydroxyurea and nocodazole, light microscopy and qRT-PCR analysis showed that, compared with unsynchronized cells and S and G2/M phase cells, SeMNPV-infected P8-Se301-C1 cells in G1 phase induced G2/M phase arrest, and the amount of virus adsorption and intracellular viral DNA replication were significantly increased (p < 0.05). In addition, budded virus (BV) production and occlusion body (OB)-containing cells were both increased at 120 h post-infection (p < 0.05). The expression of Cyclin B and CDK1 was significantly down-regulated at 48 h post-infection (p < 0.05). Finally, the arrest of SeMNPV-infected G1 phase cells in the G2/M phase increased BV production (p < 0.05) and the number of OB-containing cells. In conclusion, G1 phase infection and G2/M arrest are favorable to SeMNPV proliferation in P8-Se301-C1 cells, thereby alleviating the homologous superinfection exclusion. The results contribute to a better understanding of the relationship between baculoviruses and insect cell cycle progression and regulation.

Co-prevalence of Human Parvovirus 4 and Primate Erythroparvovirus 1 (B19) in Sickle Cell Disease and Healthy Populations

Background: Human Parvovirus 4 (P4) is a non-enveloped, single-stranded DNA virus belonging to the Tetraparvovirus genus within the Parvoviridae family. Epidemiologically, P4 exhibits similarities with its well-established family member, primate erythroparvovirus 1 (known as B19), a blood-borne virus implicated in causing aplastic crises in individuals with sickle cell disease (SCD). Despite the acknowledged association with B19, there is a dearth of prior investigations into the prevalence and clinical significance of P4 in SCD patients. Objectives: This study aims to ascertain the prevalence and outcomes of P4, along with exploring the co-prevalence of P4 with B19 in individuals with SCD. Methods: A total of 162 participants were enrolled, comprising 120 individuals with SCD and 45 healthy controls. The prevalence of P4 and B19 DNA was determined utilizing a nested-PCR method. Sequencing was performed on positive samples to validate the diagnosis, and a phylogenetic tree was constructed based on the sequencing results. Correlations in the data were analyzed using statistical methods. Results: The prevalence of P4 and B19, as well as the co-prevalence of these viruses, was significantly higher in SCD patients than in healthy individuals. Moreover, the prevalence of P4 did not exhibit a significant correlation with variables such as age, sex, aplastic crises, or specific complications associated with SCD. Conclusions: Sickle cell disease patients represent a susceptible population for P4 infection, as indicated by the heightened prevalence observed in this study.

PSI-5 Evaluation of EhV (surrogate for African swine fever virus) inactivation in corn- and soybean-based feed ingredients and diets at various times and temperatures during storage

Abstract African swine fever virus (ASFV) is a member of the nucleocytoplasmic large DNA viruses (NCLDVs) and remains stable in a variety of environments, including animal feed ingredients as shown in previous laboratory experiments and simulations. Emiliania huxleyi virus (EhV) is another member of NCLDV, which has a restricted host range limited to a species of marine algae called Emiliania huxleyi. This algal NCLDV has many similar morphological and physical characteristics to ASFV which makes it a safe and reliable surrogate for studying ASFV survival and inactivation in feed ingredient matrices in-situ under various time and temperature storage conditions. The objective of the present study was to investigate the inactivation of ASFV in various feedstuffs under a wide range of time and temperature conditions during storage using EhV as a surrogate. Corn- and soybean-based feed ingredients including corn, corn fermented protein, distiller dried grains with solubles, extruded soybean meal, high protein distiller’s grains, solvent-extracted soybean meal, soybean hulls, complete feed, and a positive control (no matrix) were inoculated with EhV (1 × 108 viruses/mL). Negative controls were inoculated with medium containing no EhV for each matrix. To assess the effects of different times and temperatures of storage, all samples were stored at temperatures of 4, 24, and 34 °C for 1, 5, 60, and 120 d. For each timepoint and temperature combination, samples were evaluated for virus presence (qPCR) and viability using a previously validated viability qPCR (v-qPCR) method. On average, we observed a 0.2 log reduction in viable viral DNA content over the 120-d period, with a 0.1 log reduction across varying temperatures. No reductions in EhV viability exceeding 1 log were detected for any matrix and the positive control at any time and temperature combination evaluated. No EhV was recovered from the negative control samples. Given that the experimental error was calculated to be within the range of 1 log, we concluded that storing EhV at 4, 24, or 34 °C for up to 120 d did not result in the inactivation of EhV in any of the ingredients, complete feed, or the control with no matrix. These results demonstrate that ASFV-like NCLDVs is extremely resilient and can maintain viability in various feed matrices during long-term storage and suggest that extending storage time alone may not be an effective mitigation practice against African swine fever virus.

Presence of Epstein-Barr virus in cerebrospinal fluid is associated with increased mortality in HIV-negative cryptococcal meningitis.

Cryptococcus neoformans is the most common cause of fungal meningitis and is associated with a high mortality. The clinical significance of concurrent Epstein-Barr virus (EBV) in the cerebrospinal fluid (CSF) of human immunodeficiency virus (HIV)-negative patients with cryptococcal meningitis (CM) remains unclear. A retrospective cohort study was performed by analyzing CSF samples from 79 HIV-negative Chinese Han patients with confirmed CM. We identified CSF viral DNA in these patients by metagenomic next-generation sequencing (mNGS) and compared 10-week survival rates among those with and without EBV DNA in CSF. Of the 79 CSF samples tested, 44.3% (35/79) had detectable viral DNA in CSF, while 55.7% (44/79) were virus-negative. The most frequent viral pathogen was EBV, which was detected in 22.8% (18/79) patients. The median number of CSF-EBV DNA reads was 4 reads with a range from 1 to 149 reads. The 10-week mortality rates were 22.2% (4/18) in those with positive CSF-EBV and 2.3% (1/44) in those with negative CSF-virus (hazard ratio 8.20, 95% confidence interval [CI] 1.52-81.80; P=0.014), which remained significant after a multivariate adjustment for the known risk factors of mortality (adjusted hazard ratio 8.15, 95% CI 1.14-92.87; P=0.037). mNGS can identify viruses that coexist in CSF of HIV-negative patients with CM. EBV DNA is most commonly found together with C. neoformans in CSF and its presence is associated with increased mortality in HIV-negative CM patients.

Anti-HBc antibodies and occult hepatitis B infection (OBI) among blood donors in Lomé, Togo

Aims: Among the progressive forms of hepatitis B virus (HBV) infections, the HBs antigen (HBsAg) remains undetectable in 25% of cases. There is also a form of HBV infection called occult HBV infection (OBI) characterized by the presence of viral DNA in the blood and tissues with no detectable levels of HBsAg associated with or without anti-HBc Ab, or anti-HBs Ab out of the pre-seroconversion period, which constitutes an infectious risk in blood transfusion. Methods: To determine the prevalence of anti-HBc Abs and the HBV DNA in the blood of donors recruited at the blood transfusion center (CNTS) of Lomé. This is a cross-sectional study including 967 blood donations which were seronegative to HBs antigen during the biological qualification of the donation (BQD) from July 28 to August 19, 2022. The detection of total anti-HBc Abs (IgM and IgG) was performed by BIORAD enzyme-linked immunosorbent assay (ELISA) method; the viral load measurement was carried out according to the polymerase chain reaction method by amplification of COBAS AmpliPrep/TaqMan 4800 (Roche diagnostics). Data processing and analysis were performed using EXCEL 2013 software and SPSS software. Concerning the ethical aspect, we have obtained written consent from the donors and the agreement of the bioethics committee for health research No. 052/2022/CBRS. Results: The prevalence of anti-HBc Abs was 33.82% (CI: 30.83–36.80). Viral DNA was detected in 10 cases. The prevalence of occult hepatitis B was 3.05% (10/327) in blood donors carrying anti-HBc antibodies, and 1.03% (10/967) in the blood donors population that does not carry HBsAg. Conclusion: The presence of occult hepatitis B within the population of blood donors at the blood transfusion center of Lomé constitutes a real threat to transfusion safety. The association of the viral DNA detection by PCR to the serological tests that are currently carried out during the biological qualification of the donations could be a better approach to reduce the risk of transmission of HBV by blood transfusion at the CNTS of Lomé.

Potency of a small molecule that targets the molluscum contagiosum virus processivity factor increases when conjugated to a tripeptide

We recently developed compound FC-7269 for targeting the Molluscum contagiosum virus processivity factor (mD4) and demonstrated its ability to inhibit viral processive DNA synthesis in vitro and cellular infection of an mD4-dependent virus (Antiviral Res 211, 2023,105520). However, despite a thorough medicinal chemistry campaign we were unable to generate a potent second analog as a requisite for drug development. We overcame this impasse, by conjugating a short hydrophobic trivaline peptide to FC-7269 to produce FC-TriVal-7269 which significantly increased antiviral potency and reduced cellular toxicity.

АНАЛИЗ КЛИНИЧЕСКОГО СЛУЧАЯ ВУЛЬГАРНОЙ ПУЗЫРЧАТКИ, АССОЦИИРОВАННОЙ С ХРОНИЧЕСКОЙ ВЭБ-ИНФЕКЦИЕЙ (ГЛОССИТА, ОБУСЛОВЛЕННОГО ВЭБ)

Background. One of the autoimmune diseases is pemphigus, a chronic malignantly occurring autoimmune dermatosis accompanied by the development of blisters on the skin and mucous membranes. Isolated lesions of the SOPR occur in 10–20% of cases, complicating differential diagnosis. Numerous studies have proven the role of herpes virus infection as an aggravating factor in the course of true acantholytic pemphigus. The aim of the study was to determine the features of the clinical course of vulgar pemphigus against the background of reactivation of chronic EBV infection (glossitis caused by EBV). The clinical case of a patient with a diagnosis of “Vulgar pemphigus. Glossitis caused by the Epstein-Barr virus”. The anamnesis of the disease was carefully collected, a clinical and laboratory examination was performed with a cytological examination of a smear taken from a LITTER, a general blood test with an expanded leukocyte formula, C-reactive protein. In order to diagnose the viral load, PCR diagnostics was performed on the DNA of the herpes virus 1, 2, 4, 5 types in saliva, on the DNA of EBV in the blood, quantitatively. Specific antibodies of Ig classes M and G to EBV were determined by the ELISA method. Results. Due to the lifelong persistence of EBV in the structures of the oral cavity, its reactivation leads not only to chronic pastoral infection and the local development of the corresponding glossitis, but also, first of all, to a decrease in the level of local immunity. This fact aggravates the course of pemphigus, shortens the period of absence of relapse. The appointment of antiviral therapy at the first visit allowed to stabilize the symptoms caused by EBV, which led to positive dynamics in the course of pemphigus against the background of average dosages of prednisone in a shorter time. Conclusions. With clinical and laboratory confirmation of chronic EBV infection (glossitis caused by EBV), it is recommended to include antiviral and immunocorrective drugs, systemically and topically, in the complex treatment of pemphigus erythematosus.

Epstein-Barr Virus in the Cerebrospinal Fluid and Blood Compartments of Patients With Multiple Sclerosis and Controls.

Epstein-Barr virus (EBV) infection is a major risk factor of multiple sclerosis (MS). We examined the presence of EBV DNA in the CSF and blood of patients with MS and controls. We analyzed whether EBV DNA is more common in the CSF of patients with MS than in controls and estimated the proportions of EBV-positive B cells in the CSF and blood. CSF supernatants and cells were collected at diagnostic lumbar punctures from 45 patients with MS and 45 HLA-DR15 matched controls with other conditions, all participants were EBV seropositive. Cellular DNA was amplified by Phi polymerase targeting both host and viral DNA, and representative samples were obtained in 28 cases and 28 controls. Nonamplified DNA from CSF cells (14 cases, 14 controls) and blood B cells (10 cases, 10 controls) were analyzed in a subset of participants. Multiple droplet digital PCR (ddPCR) runs were performed per sample to assess the cumulative EBV positivity rate. To detect viral RNA as a sign of activation, RNA sequencing was performed in blood CD4-positive, CD8-positive, and CD19-positive cells from 21 patients with MS and 3 controls. One of the 45 patients with MS and none of the 45 controls were positive for EBV DNA in CSF supernatants (1 mL). CSF cellular DNA was analyzed in 8 independent ddPCRs: EBV DNA was detected at least once in 18 (64%) of the 28 patients with MS and in 15 (54%) of the 28 controls (p = 0.59, Fisher test). The cumulative EBV positivity increased steadily up to 59% in the successive ddPCRs, suggesting that all individuals would have reached EBV positivity in the CSF cells, if more DNA would have been analyzed. The estimated proportion of EBV-positive B cells was >1/10,000 in both the CSF and blood. We did not detect viral RNA, except from endogenous retroviruses, in the blood lymphocyte subpopulations. EBV-DNA is equally detectable in the CSF cells of both patients with MS and controls with ddPCR, and the probabilistic approach indicates that the true positivity rate approaches 100% in EBV-positive individuals. The proportion of EBV-positive B cells seems higher than previously estimated.

Phenotypic assessment of antiviral activity for spiro-annulated oxepanes and azepenes.

Evolutionary potential of viruses can result in outbreaks of well-known viruses and emergence of novel ones. Pharmacological methods of intervening the reproduction of various less popular, but not less important viruses are not available, as well as the spectrum of antiviral activity for most known compounds. In the framework of chemical biology paradigm, characterization of antiviral activity spectrum of new compounds allows to extend the antiviral chemical space and provides new important structure-activity relationships for data-driven drug discovery. Here we present a primary assessment of antiviral activity of spiro-annulated derivatives of seven-membered heterocycles, oxepane and azepane, in phenotypic assays against viruses with different genomes, virion structures, and genome realization schemes: orthoflavivirus (tick-borne encephalitis virus, TBEV), enteroviruses (poliovirus, enterovirus A71, echovirus 30), adenovirus (human adenovirus C5), hantavirus (Puumala virus). Hit compounds inhibited reproduction of adenovirus C5, the only DNA virus in the studied set, in the yield reduction assay, and did not inhibit reproduction of RNA viruses.

Microhomology-mediated repair machinery and its relationship with HPV-mediated oncogenesis.

Human Papillomaviruses (HPV) are a diverse family of non-enveloped dsDNA viruses that infect the skin and mucosal epithelia. Persistent HPV infections can lead to cancer frequently involving integration of the virus into the host genome, leading to sustained oncogene expression and loss of capsid and genome maintenance proteins. Microhomology-mediated double-strand break repair, a DNA double-stranded breaks repair pathway present in many organisms, was initially thought to be a backup but it's now seen as vital, especially in homologous recombination-deficient contexts. Increasing evidence has identified microhomology (MH) near HPV integration junctions, suggesting MH-mediated repair pathways drive integration. In this comprehensive review, we present a detailed summary of both the mechanisms underlying MH-mediated repair and the evidence for its involvement in HPV integration in cancer. Lastly, we highlight the involvement of these processes in the integration of other DNA viruses and the broader implications on virus lifecycles and host innate immune response.

Hepatitis B virus DNA is absent in Lebanese blood donors who are positive for both anti-HBC and anti-HBS antibodies

Hepatitis B virus DNA is absent in Lebanese blood donors who are positive for both anti-HBC and anti-HBS antibodies

2039: Validation and Utility of Circulating Tumor Tissue Modified Viral HPV DNA in a Community HN-MDC

2039: Validation and Utility of Circulating Tumor Tissue Modified Viral HPV DNA in a Community HN-MDC

The role of IFI16 in regulating PANoptosis and implication in heart diseases.

Interferon Gamma Inducible Protein 16 (IFI16) belongs to the HIN-200 protein family and is pivotal in immunological responses. Serving as a DNA sensor, IFI16 identifies viral and aberrant DNA, triggering immune and inflammatory responses. It is implicated in diverse cellular death mechanisms, such as pyroptosis, apoptosis, and necroptosis. Notably, these processes are integral to the emergent concept of PANoptosis, which encompasses cellular demise and inflammatory pathways. Current research implies a significant regulatory role for IFI16 in PANoptosis, particularly regarding cardiac pathologies. This review delves into the complex interplay between IFI16 and PANoptosis in heart diseases, including atherosclerosis, myocardial infarction, heart failure, and diabetic cardiomyopathy. It synthesizes evidence of IFI16's impact on PANoptosis, with the intention of providing novel insights for therapeutic strategies targeting heart diseases.

Activation of the cGAS-STING pathway by viral dsDNA leading to M1 polarization of macrophages mediates antiviral activity against hepatitis B virus

Background and aimsActivation of the cGAS-STING pathway induces the production of type I interferons, initiating the antiviral immune response, which contributes to the clearance of pathogens. Previous studies have shown that STING agonists promote hepatitis B virus (HBV) clearance; however, few studies have investigated the effect of activating the cGAS-STING pathway in macrophages on HBV. MethodsThe polarization status of HBV particle-stimulated RAW264.7 macrophages was analyzed. After stimulation with HBV particles, the analysis focused on determining whether the DNA sensors in RAW264.7 macrophages recognized the viral double-stranded DNA (dsDNA) and evaluating the activation of the cGAS-STING pathway. Coculture of mouse macrophages and hepatocytes harboring HBV was used to study the antiviral activity of HBV-stimulated RAW264.7 macrophages. ResultsAfter stimulation with HBV particles, HBV relaxed circular DNA (rcDNA) was detected in RAW264.7 macrophages, and the protein expression of phospho-STING, phospho-TBK1, and phospho-IRF3 in the STING pathway was increased, as shown by Western blot analysis, which revealed that M1 polarization of macrophages was caused by increased expression of CD86. RT–PCR analyses revealed elevated expression of M1 macrophage polarization-associated cytokines such as TNFα, IL-1β, iNOS, and IFNα/β. In the coculture experiment, both HBsAg and HBeAg expression levels were significantly decreased in AML12-HBV1.3 cells cocultured with the supernatants of HBV-stimulated RAW264.7 macrophages. ConclusionThe results suggest that macrophages can endocytose HBV particles. Additionally, viral dsDNA can be recognized by DNA pattern recognition receptors, which in turn activate the cGAS-STING pathway, promoting the M1 polarization of macrophages, while no significant M2 polarization is observed. Macrophages stimulated with HBV particles exhibit enhanced antiviral activity against HBV.

Significance of Pre-Treatment CALLY Score Combined with EBV-DNA Levels for Prognostication in Non-Metastatic Nasopharyngeal Cancer Patients: A Clinical Perspective.

The C-reactive protein-albumin-lymphocyte (CALLY) score is a novel indicator associated with inflammation, immunity, and nutrition, utilized for cancer prognostic stratification. This study aimed to evaluate the integrated prognostic significance of the pre-treatment CALLY score and Epstein-Barr virus (EBV) DNA levels in nasopharyngeal carcinoma (NPC) patients and to develop prognostic models. A total of 1707 NPC patients from September 2015 to December 2017 were retrospectively enrolled. The cut-off point for the CALLY score, determined by maximum selected rank statistics, integrates with the published cut-off point for pre-EBV DNA to develop a comprehensive index. Subsequently, patients were randomly allocated in a 1:1 ratio into training and validation cohorts. Survival analysis was conducted using the Kaplan-Meier method with Log rank tests, and the Cox proportional hazards model was applied to identify independent prognostic factors for constructing predictive nomograms. The predictive ability of the nomograms were assessed through the concordance index (C-index), calibration curves, and decision curve analysis. By integrating CALLY scores and EBV-DNA levels, patients were categorized into three risk clusters. Kaplan-Meier curves reveal significant differences in overall survival (OS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) outcomes among different risk groups (all P values < 0.05). Multivariate analysis revealed that CALLY-EBV DNA index serves as an independent prognostic factor for the OS, DMFS, and LRRFS. The prognostic nomograms based on the CALLY-EBV DNA index provided accurate predictions for 1-year, 3-year, and 5-year OS, DMFS, and LRRFS. Additionally, compared to the traditional TNM staging system, the nomograms exhibited enhanced discriminatory power, calibration capability, and clinical applicability. All results were in agreement with the validation cohort. The CALLY-EBV DNA index is an independent prognostic biomarker. The nomogram prediction models, constructed based on the CALLY-EBV DNA index, demonstrates superior predictive performance compared to the traditional TNM staging.