Since mid-May 2026, a rapidly evolving Ebola virus disease (EVD) outbreak caused by the Bundibugyo strain (Orthoebolavirus bundibugyoense) has been spreading in eastern Democratic Republic of Congo (Ituri, North and South Kivu provinces) and Uganda (Kampala and Wakiso areas). Uganda has so far reported 14 imported cases and 5 secondary transmissions according to the World Health Organization (WHO). As of 30 July 2026, 3605 confirmed cases, including 1587 deaths, have been reported in the Democratic Republic of the Congo. Unlike previous Zaire outbreaks, no specific licensed vaccine or antiviral treatment exists for the Bundibugyo strain, making behavioral and community-based interventions the only line of defense. However, armed conflict, chronic insecurity, massive population displacements (>2.1 million), and profound community distrust of health authorities are crippling the response. This article therefore argues that the current threat is a major warning sign for the Great Lakes region, and that investing in Risk Communication and Community Engagement (RCCE) such as involving community health workers, natives, and trusted leaders is the most urgent priority to avoid a regional health catastrophe. Without a humanitarian truce to guarantee access and a coordinated cross-border RCCE strategy, undetected transmission will continue to fuel this outbreak and future zoonotic emergencies.
We performed a population-based study in densely populated areas of the largest city of Pakistan (Karachi) to determine the overall type- and age-specific prevalence of human papillomavirus (HPV) in the setting of an unscreened and non-vaccinated female population between May 2022 and November 2023. Women (n=3,119) were invited to participate in the study from whom a total of 497 women gave consent and provided cervical samples. HPV positivity was determined and specific HPV genotypes were identified using the INNO LiPa Extra II-line probe assay. Total HPV positivity among all age groups was 16.7%. High risk HPV types (groups 1 and 2) were found in 11.9% whereas low risk types and unclassified types were 4.8% in all samples. Fourteen HR types were detected as single infections (8.4%), highest prevalent types are HPV-31 and HPV-53 (each 1.2%) followed by HPV-68 (1.0%), HPV-16 (0.8%), HPV-33, -39 and -51 (each 0.6%). While HPV-35, HPV-52, HPV-70, HPV-73 and HPV-82 were the least prevalent types. The most prevalent (1.2%) low-risk HPV type detected was HPV-6 of all samples. The highest HPV prevalence (21.3%) was observed in subjects aged 25–34 years (n=220), whereas in the age group above 54 years (n=24) we detected HPV in 12.5% of samples tested. Single HR HPV infections were observed in 8.4%, while multiple HR infections were detected in 4.02% of the 497 women tested. Extrapolating this data to the total female population of Pakistan allows to estimate that about 20 million women are HPV positive. This rough estimation forms a strong basis of an organized cervical cancer screening program using high precision HPV tests for early detection of HPV infections and related diseases including cervical cancer. In addition, this also establish the need of implementation of immunization program with the recently licensed nonavalent human papillomavirus vaccine. This can significantly reduce the future morbidity and mortality from cervical and oral cancer, pre-cancerous lesions and other HPV-related cancers in the female and male population of Pakistan. The current study, therefore, provide a credible basis of further research, and follow-up action on HPV-related disease burden.
Background:The incidence of low-level viremia (LLV) in chronic hepatitis B (CHB) patients receiving first-line nucleos(t)ide analogue (NAs) therapy with negative results on conventional HBV DNA testing is unknown. Methods:A total of 557 treatment-naïve CHB patients who had received first-line NAs therapy for more than 48 weeks and had negative results on conventional HBV DNA testing were enrolled. According to the results of two high-sensitivity HBV DNA tests, CHB patients were divided into two categories: maintained virological response (MVR) and LLV. Finally, LLV patients were divided into two groups: maintaining NAs therapy and switching NAs therapy. Results:Among the patients whose conventional HBV DNA testing was negative, the proportions of MVR and LLV were 64.1% and 35.9% respectively. Age and treatment duration are negatively correlated with the occurrence of LLV, while HBsAg level, HBeAg positivity, and ALT level were positively associated with the occurrence of LLV. At the 48th week of follow-up, the complete virological response (CVR) of the switching NAs group and the maintaining NAs group were 81.63% and 86.36% respectively (P = 0.536). In addition, there were no statistically significant differences between the two groups in the HBeAg-positive rate, HBsAg level, ALT level, APRI, and FIB-4 score over the 48-week follow-up period. Conclusion:In CHB patients who had negative results of conventional HBV DNA testing, the incidence rate of LLV is still high. For LLV patients with negative results in conventional HBV DNA testing, the vast majority of patients can achieve CVR whether maintaining or switching NAs, and the efficacy of the two treatment strategies is similar. However, these findings may not be applicable to patients with higher baseline HBV DNA levels, different distributions of NAs types, or those treated in other clinical settings.
Background:Elderly patients remain at high risk for adverse outcomes from COVID-19 despite the generally reduced pathogenicity of the Omicron variant. Corticosteroids are commonly used in hospitalized patients with COVID-19, particularly in those with more severe disease, while antiviral agents inhibit viral replication. However, evidence comparing corticosteroid monotherapy with corticosteroid-antiviral combination therapy in elderly hospitalized patients remains limited. Methods:This multicenter retrospective cohort study included hospitalized patients aged ≥60 years with laboratory-confirmed COVID-19 at two tertiary hospitals in China, all of whom received systemic corticosteroid therapy during hospitalization. Patients were classified into either a corticosteroid monotherapy group or corticosteroid-antiviral combination therapy group. To reduce confounding, inverse probability of treatment weighting (IPTW) based on propensity scores was applied. The primary outcome was all-cause in-hospital mortality, and mechanical ventilation was assessed as a secondary outcome. Results:A total of 624 elderly hospitalized patients with COVID-19 who received systemic corticosteroid therapy were included in the IPTW analysis (290 in the corticosteroid group and 334 in the combination therapy group). After weighting, baseline characteristics were well balanced between groups. The weighted incidence of in-hospital mortality was lower in the combination therapy group than in the corticosteroid monotherapy group (8.09% vs 13.47%; OR 0.566, 95% CI 0.333-0.961; p = 0.035). No statistically significant difference was observed in the risk of mechanical ventilation between groups (OR 1.286, 95% CI 0.924-1.789; p = 0.136). Conclusions:Among elderly hospitalized patients with COVID-19 during the Omicron period who received systemic corticosteroid therapy corticosteroids combined with antiviral therapy were associated with lower in-hospital mortality compared with corticosteroid monotherapy, while no significant difference was observed in mechanical ventilation.
The transition from traditional biomedical-based vaccination campaigns in Pakistan has faced many difficulties. This paper proposes that the central problem is not the content of vaccine-related messages per se, but whether their targets perceive those messages as aligned with their identities. The objective of this study is to examine the effects of message framing and perceived identity threat on polio intention to vaccinate through the theoretical perspectives of Framing Theory and Social Identity Theory (SIT). A survey sample of 600 adults in Sindh and Balochistan, two regions understudied in empirical research on this topic, was employed. Hierarchical multiple regression and moderation analyses were used to test the hypotheses. Collective identity framing was the most significant positive predictor of intention to vaccinate (β = .41, p < .001). Identity threat, the perception that an anti-polio message contradicts the respondents' religious or cultural identity, was the strongest negative predictor (β = -.45, p < .001). Peer-norm salience significantly mediated the link between framing uptake and collective framing among community-oriented respondents. Critically, religious commitment was non-significant once identity threat was controlled, demonstrating that identity threat, not religiosity, is the proximal driver of refusal. The Identity-Anchored Message Uptake (IAMU) Model is proposed based on these findings.
Chikungunya virus (CHIKV) is a re-emerging arbovirus causing acute febrile illness and chronic debilitating arthritis, thereby imposing significant global public health and economic burdens. While robust immune responses involving inflammatory cytokines and immune cell infiltration characterize acute infection, the cellular mechanisms underlying pathogenesis and chronicity remain incompletely defined. In this study, we employed single-cell RNA sequencing (scRNA-seq) to comprehensively profile splenic and peripheral blood mononuclear cells (PBMCs) immune responses in rhesus macaques at day 7 post CHIKV infection, an acute phase. Splenic neutrophils marked recruitment, upregulation of S100A8/S100A9, downregulation of interferon-stimulated genes (ISGs), and functional activation marked by degranulation, enhanced anti-apoptotic pathways, and neutrophil extracellular traps (NETs) formation, as visualized by multiplex immunofluorescence. Pseudotime trajectories delineated progressive states transitioning from proliferation to effector functions. Splenic T and B cells showed increased abundance with innate-to-adaptive transition signature. Splenic CD4+ and CD8+ T cell subsets and B cell subsets exhibited enrichment for innate immune pathways and translational machinery. In PBMCs, CD8+ T cell subsets and B cell subsets exhibited activation of adhesion, cytokine signaling, and protein homeostasis pathways in infection group. Notably, CHIKV infection induced skewing of T cells toward states enriched for Th1 and Th17 differentiation (marked by NXPE3, LEF1, NFKBIZ expression) in PBMCs. These results delineate the spatiotemporal immune landscape during CHIKV acute phase, in this non-human primate model, identifying neutrophil-mediated inflammation, lymphocyte transcriptional adaptation, and Th1/Th17 polarization as hallmarks of acute phase, offering a detailed cellular map that may inform future investigations into CHIKV pathogenesis and therapeutic strategies.
Background:This study aims to describe the epidemiological characteristics of human rabies cases in Côte d'Ivoire over 11-year. Methods:A retrospective analysis was conducted on all human rabies cases reported from 2013 to 2023. Samples were analyzed by RT-qPCR using primers targeting the nucleoprotein gene according to WHO rabies diagnostic guidelines. Sociodemographic, clinical, and epidemiological data were compiled and analyzed. Results:Hundred and fifty-two (84.44 %) out of hundred and eighty suspected cases were positive, representing an annual average of 14 cases of rabies. Men accounted for 2/3 of suspected cases and 68.42 % of confirmed ones. Patients' average age was 25.98 years. Socio-professional groups most affected were schoolchildren with 28.29 % of cases, farmers (20.39 %), and housekeepers (13.16 %). District of Abidjan, along with Loh-Djiboua, Gbêkê and Haut-Sassandra regions, recorded the highest positivity rates. Most infections (90.8 %) resulted from third-degree contact (bites), mainly to the upper limbs (46.71 %). The spastic form of rabies was predominant (84.87 %). None of the positive patients had been vaccinated. Vectors were mainly wandering dogs (82.24 %). Conclusion:Human rabies remains a major public health concern in Côte d'Ivoire, mainly affecting schoolchildren and mostly located in rural areas. Prevention strategies, including vaccination and control of wandering animals, remain essential.
Persistent infection with high-risk human papillomavirus (HR HPV) is the necessary cause of cervical cancer. In the Philippines, available data on the prevalence and genotype distribution of HPV infections are limited and largely derived from earlier hospital-based studies. The present study determined the overall and type-specific prevalence of HR HPV infection among women in selected communities in the Philippines, along with the associated sociodemographic and behavioral factors. A total of 1,194 women from two communities were examined. Cervical swabs were collected, and the extracted DNA were analyzed for HPV genotyping through a commercial multiplex real-time PCR assay kit. Sociodemographic information, clinical history, and sexual and reproductive behavior were obtained through an interviewer-administered semi-structured questionnaire. The overall prevalence of HR HPV infection was 11.22 % (95 % CI: 9.56-13.14 %). Women residing in urban areas had 1.62 times higher odds (95 % CI: 1.08-2.42) of HR HPV infection than those in rural areas. Moreover, for every year of delaying vaginal sexual debut, there was a 7 % decrease in the odds of HR HPV infection. HPV 52 was the most prevalent genotype (2.59 %), followed by HPV 16 (1.84 %) and HPV 68 (1.09 %). Multiple HR HPV genotypes were recorded in 23 % of HR HPV-infected women. The most frequent co-infections are HPV 16 + HPV 18, HPV 16 + HPV 52, and HPV 39 + HPV 52. These findings highlight the need for updated surveillance and consideration of local genotype distribution in cervical cancer prevention strategies, such as in HPV DNA testing and HPV vaccination programs.
Background:Hepatitis B virus (HBV) infection is a major risk factor for liver fibrosis, cirrhosis, and hepatocellular carcinoma. Whether patients with chronic hepatitis B (CHB) receiving oral nucleos(t)ide analogue (NA) therapy can safely discontinue treatment after HBsAg seroclearance is attracting clinical attention.This study aimed to explore the maintenance rate of HBsAg-negative status and the predictors of HBsAg repositivity after drug withdrawal in non-cirrhotic CHB patients who had achieved HBsAg seroclearance following long-term NAs therapy. Methods:This is a single center retrospective study focusing on non-cirrhotic CHB patients who received NAs treatment and achieved HBsAg seroclearance.These patients were treated in the hepatitis clinic of West China Hospital of Sichuan University and followed up for a long time. CHB patients were required to have complete demographic and clinical data. Additionally, serum levels of HBV RNA and HBcrAg were measured in all patients at the time of NAs cessation. Results:A total of 137 non-cirrhotic CHB patients with HBsAg seroclearance were screened. Ultimately, 54 patients agreed to discontinue treatment, while 83 declined. Among the 54 patients who terminated treatment, 43 were male and 11 were female. Of these discontinued patients, 44 received continuous monotherapy, while 10 received combination therapy. All patients in this study received NAs antiviral treatment for more than 5 years. 79.6 % (43/54) of patients were found to be positive for HBsAb and 59.3 % (32/54) of patients had HBsAb≥200 IU/ml at the time of NAs discontinuation. Among the discontinued patients, all 54 patients were HBV RNA negative, and 87.0 % (47/54) were HBcrAg negative.The rates of HBsAg repositive were 3.7 % (95 % CI, 0.6 %-12.7 %) and 9.3 % (95 % CI, 3.8 %-19.7 %) at 24 and 48 weeks after drug withdrawal, respectively, and 3 of them were accompanied by HBV DNA relapse. All patients who regained HBsAg positivity after NAs discontinuation had serum HBcrAg levels greater than 3 log10 U/mL at the time of discontinuation. Conclusion:In this single-center cohort, most non-cirrhotic patients who achieved HBsAg seroclearance on long-term NAs therapy maintained HBsAg loss over 48 weeks after discontinuation. HBcrAg positivity at end of treatment was observed in all cases of HBsAg reappearance, suggesting HBcrAg may help identify patients at higher short-term risk of seroreversion. Larger, longer-term studies are required to confirm these findings.
Background and objectives:Pegylated interferon (PegIFN) is one of the few strategies capable of achieving hepatitis B surface antigen (HBsAg) seroclearance in chronic hepatitis B (CHB) patients. However, its role in HBeAg-negative chronic HBV infection (also referred to as immune-inactive or "inactive carrier" phase) remains unclear. This study compared PegIFN efficacy between immune-inactive and immune-active patients and explored predictors of treatment response. Research methods:We retrospectively analyzed 211 consecutive HBeAg-negative patients treated with PegIFN α-2b in our center from 2019 to 2023, including 139 with immune-inactive chronic HBV infection and 72 immune-active CHB patients. Effectiveness was assessed in the full analysis set (FAS) and efficacy in the per-protocol set (PPS). Predictors of HBsAg seroclearance were identified using Cox regression. Results:At week 48, overall HBsAg seroclearance was 26.5 % (FAS), with comparable outcomes between immune-inactive and immune-active groups (26.6 % vs. 26.4 %, P > 0.05). Inactive patients receiving PegIFN + nucleos(t)ide analogues (NAs) showed no added benefit. In the PPS, clearance reached 69.1 %, but 40.3 % discontinued prematurely-mostly due to subjective intolerance-highlighting a major gap between real-world effectiveness and theoretical efficacy. Multivariate analysis identified two independent predictors: baseline HBsAg <100 IU/mL (HR 2.999, 95 % CI 1.625-5.536, P < 0.001) and on-treatment HBsAg decline ≥1 log10 IU/mL within any 12-week interval (HR 11.205, 95 % CI 4.379-28.674, P < 0.001). Conclusions:PegIFN α-2b achieved clinically meaningful HBsAg seroclearance in both immune-inactive and immune-active patients. Early discontinuation markedly reduced real-world effectiveness. Baseline low HBsAg and dynamic on-treatment decline are pragmatic predictors for optimizing patient selection and guiding interferon-based strategies.
The presence of neutralizing antibodies is considered a surrogate marker of protection against the three serotypes of poliovirus. The need to use the Microneutralization test in assessing the neutralizing antibodies to the three serotypes of polioviruses among vaccinated children aged 1-15 years informed this study. Of 400 children tested, 309 (77.3 %), 253 (63.3 %), and 308 (77.0 %) had neutralizing antibodies against P1, P2, and P3, respectively. Only 191 (47.8 %) had neutralizing antibodies against P1P2P3 simultaneously, the global target. Whilst 91 (22.8 %) had no neutralizing antibodies against P1, these children were protected against P2 (23.0 %) and P3 (43.9 %). Similarly, 147 (36.8 %) children had no neutralizing antibodies against P2, but were protected against P1 (66.0 %) and P3 (65.3 %). Furthermore, 52(13 %), 51(12.8 %), and 52 (13.0 %) had no neutralizing antibodies against the combination of P1P3, P1P2, and P2P3, respectively. Only 34 (8.5 %) of the children had no nAb to any of the three serotypes. The optimal number of Polio vaccine doses for effective immunity varied depending on the serotype. Also, gender differences may favor the speed at which children achieve the target antibody titers. Higher antibody titers (1:1280) were observed for P2 and P3, with six of the children having a titer of 1:10240 for P3. The combination of supplementary immunization activities and routine immunization generated a robust immune response across all poliovirus serotypes, in contrast to each of the two. The administrative data and population immunity were not commensurate. New strategies to increase immunity against the P1P2P3 simultaneously in all age groups are urgently required.
Background:The HIV/AIDS epidemic remains a critical public health challenge in Bangladesh, with complex epidemiological trends and sex-specific disparities requiring detailed investigation to guide effective interventions. This study comprehensively analyzes the temporal dynamics of HIV/AIDS burden from 1990 to 2021 and employs advanced statistical modeling to forecast future trends up to 2050, aiming to inform targeted public health strategies. Methods:Data on Disability-Adjusted Life Years (DALYs), deaths, incidence, prevalence, Years Lived with Disability (YLDs), and Years of Life Lost (YLLs) were sourced from a robust epidemiological database for Bangladesh. Joinpoint regression analysis was utilized to detect significant trend changes, calculating the average annual percent change (AAPC) and annual percent change (APC) with 95 % confidence intervals (CIs). Sex-stratified analyses elucidated disparities between males and females. Autoregressive Integrated Moving Average (ARIMA) models were applied to project age-standardized rates (ASRs) for each metric through 2050, incorporating historical trends and variability to ensure robust predictions. Results:From 1990 to 2021, the HIV/AIDS burden in Bangladesh increased significantly across all metrics, with distinct sex-specific patterns. The overall AAPC for DALYs was 19.0332 % (95 % CI: 15.8145, 23.0026, p < 0.000001), with females showing a higher AAPC (21.7252 %, 95 % CI: 18.2308, 25.7026) than males (18.4703 %, 95 % CI: 15.1896, 22.5646). Deaths exhibited a similar trend, with an overall AAPC of 18.9645 % (95 % CI: 15.7008, 23.2166), higher in females (21.8808 %, 95 % CI: 18.4195, 25.9932) than males (18.4655 %, 95 % CI: 15.1992, 22.6832). Incidence rose with an AAPC of 15.7929 % (95 % CI: 11.6467, 19.1922), slightly higher in females (16.2581 %, 95 % CI: 12.1795, 20.0487) than males (15.1639 %, 95 % CI: 11.2952, 18.4817). Prevalence increased markedly (AAPC: 18.2239 %, 95 % CI: 14.5688, 22.2078), with females at 20.2887 % (95 % CI: 16.6453, 24.3006) and males at 18.1837 % (95 % CI: 14.6734, 21.841). YLDs and YLLs followed similar patterns, with females consistently showing higher AAPCs. Joinpoint analysis identified peak APCs in the 1990s and early 2000s, followed by moderated growth and declines post-2016, particularly from 2019 to 2021, reflecting potential intervention impacts. ARIMA forecasts project a decline in DALYs, deaths, and YLLs ASRs to negligible levels by 2050 for both sexes, with wide CIs indicating substantial uncertainty (e.g., DALYs overall: negligible, 95 % CI: 0, 583.3697). Incidence ASRs are expected to stabilize (e.g., overall: 0.963899, 95 % CI: 0, 3.819279), while prevalence ASRs are projected to rise dramatically, particularly for males (614.7463, 95 % CI: 0, 695680.4), highlighting significant long-term challenges. Conclusions:The HIV/AIDS burden in Bangladesh has escalated significantly from 1990 to 2021, with females bearing a disproportionately higher burden across all metrics. Recent declines suggest the efficacy of public health interventions, but persistent sex disparities and projected prevalence increases underscore the need for targeted, sex-specific strategies. The considerable uncertainty in long-term forecasts emphasizes the importance of sustained surveillance, adaptive interventions, and resource allocation to mitigate the epidemic's impact by 2050, ensuring equitable health outcomes.
The persistence of latently infected CD4+ T cells is the major barrier to cure of people with HIV (PWH) on antiretroviral therapy (ART). While most latently infected cells are transcriptionally silent, some express low levels of cell associated (CA) HIV RNA. In this prospective controlled interventional study, we tested the hypothesis that acute psychological stress could drive HIV transcription in PWH on ART. PWH on suppressive ART underwent the Trier Social Stress Test (TSST) and comparisons were made to a similar period of time without an intervention (control). During the test, physiological markers of acute psychological stress including pre-ejection period and cardiac output changed in all participants, as anticipated. Compared to the control day, the TSST led to a significant increase in CA HIV RNA with no change in the level of cell associated HIV DNA, indicating an increase in HIV transcription in response to stress. Change in HIV transcription was associated with physiological markers of stress but not with changes in immune cells. These data demonstrate that HIV transcription is increased following acute stress and have implications on the impact of stress on the HIV reservoir and the design of cure strategies for PWH.
Introduction:Directly observed therapy (DOT) is an effective strategy to optimize hepatitis C virus (HCV) cure rates in people who inject drugs (PWID) on stable opioid agonist therapy (OAT). While adherence to daily DOT is excellent, it remains unclear if extended DOT distribution intervals result in similar sustained virologic response (SVR) rates. Methods:PWID undergoing DOT with direct-acting antiviral agents (DAA) alongside OAT for HCV infection at a low-threshold institution were included. Social distancing requirements during the COVID19 pandemic led to an extension in DOT dispensation intervals; therefore, the study population was classified according to "tight period" (DAA start 2014-2020) and "liberal period" (DAA start 2020-2023) cohorts. Socioeconomic characteristics, DOT distribution schedules, and rates of SVR at week 12 after end of therapy (SVR12) were compared between groups. Results:We included 719 consecutive PWID (male: 76.5 %; median age: 39 years), 441 (61.3 %) were treated in the "tight period" and 278 (38.7 %) in the "liberal period". Baseline characteristics were comparable between cohorts, however, socioeconomic features of the "liberal period" group showed more problematic features (unemployment: 83.1 % vs. 67.3 %; lack of housing: 38.5 % vs. 35.1 %; ongoing injection drug use: 64.0 % vs. 57.8 %; each p < 0.001).While the "tight period" group had their DAA most commonly dispensed on a daily basis (78.9 %), the "liberal period" group received their DAA/OAT mostly once weekly (45.0 %) or 2-3x/week (24.1 %; p < 0.001). The number of missed DAA ingestions (0.3 % vs. 0.4 %; p = 0.239) and SVR12 rates by modified intention to treat analysis (exclusion of PWID who were lost to follow-up [FU] or died) were similar (401/404, 99.3 % vs. 194/195, 99.5 %; p = 1.000) between tight and liberal period, respectively.Loss of FU after end of DAA treatment was more common during the "liberal period" (28.8 % vs. 7.9 %; p < 0.001), yet no treatment interruptions or early discontinuations occurred. Conclusion:DOT originally aimed to address non-adherence among PWID by strict control through daily supervised OAT and DAA ingestion. While this approach inherently manifests a position of advanced distrust towards PWID, our findings suggest that a strategy of advanced trust and liberalization of DOT alongside effective harm reduction measures yields excellent adherence and results in similarly high HCV cure rates.
Epstein-Barr virus-associated gastric carcinoma (EBVaGC), a distinct subtype of gastric cancer, accounts for approximately 10 % of all gastric cancer cases. 2-deoxyglucose (2-DG), a glycolysis inhibitor, has emerged as a crucial tool in cancer therapy. However, the differential effects of 2-DG on EBVaGC and EBV-negative gastric carcinoma (EBVnGC) are not yet fully understood. In this study, we demonstrated that 2-DG inhibited the proliferation of both AGS and AGS-EBV cells, with AGS-EBV cells exhibiting greater sensitivity, particularly under hypoxic conditions. Furthermore, EBV infection was found to upregulate glycolytic gene expression in AGS-EBV cells, particularly under hypoxic conditions, through HIF-1α-dependent mechanisms. Notably, 2-DG also inhibited EBV lytic reactivation in AGS-EBV cells under hypoxic conditions. These findings provide valuable insights into the molecular mechanisms of EBV-mediated metabolic reprogramming and highlight the potential of 2-DG as a therapeutic agent for EBVaGC.
Integrase strand transfer inhibitors (INSTIs) are the cornerstone of modern antiretroviral therapy (ART), achieving durable plasma HIV-1 suppression in most people living with HIV (PLWH). Previous comparisons of INSTI- and non-INSTI-based regimens have largely focused on HIV reservoir proviral assessments- typically total HIV DNA -without assessing reservoir activity. In this first functional comparison, we measured cell-associated (CA) short HIV-1 RNA transcripts, a marker of active transcription, alongside HIV-1 DNA in white blood cells from 92 virally suppressed individuals on INSTI-based (n = 73) or non-INSTI-based (n = 19) ART. CA short RNA transcripts were detected in all participants and HIV-1 DNA in 99 %, despite undetectable plasma viremia in >93 %. Individuals with prior "blips" - defined as a maximum of two episodes with 20-200 copies/mL plasma HIV-1 RNA over more than two years - had significantly higher CA RNA and HIV DNA than non-blip participants, confirming our previous findings. However, reservoir size and transcriptional activity did not differ significantly between INSTI and non-INSTI groups. These findings indicate that while INSTIs effectively block new integration events, they do not suppress ongoing transcription from the latent reservoir. Therapeutic strategies directly targeting HIV transcription should therefore be prioritized in cure-oriented research for PLWH on long-term suppressive ART.
The persistent challenge posed by viruses such as HIV, HBV, and SARS-CoV-2 necessitates the continuous evolution of molecular tools for their study and for advancing therapeutic research. Split-protein complementation assays (PCAs), where a reporter protein is divided into two inactive fragments, have evolved from simple reporters of biological events into an increasingly important tool in modern virology. This review traces the evolutionary trajectory of split-protein systems. We begin with their foundational use in mechanistic discovery, where they first visualized viral-host interactions in living cells. We then explore their translation into practical applications, such as high-throughput drug screening and rapid point-of-care diagnostics. A step in this evolution was the development of systematic engineering platforms, dramatically accelerating the creation of novel biosensors. Finally, we discuss the latest frontier: engineering therapeutically active ''split effectors.'' By integrating principles from synthetic biology, these advanced systems can function as programmable logic gates that respond to specific viral signatures. While therapeutic translation remains preclinical, split-protein platforms are emerging as tangible tools for advanced research and potential therapeutic development.
