Search Results (374)

The vast majority of viruses causing human and animal diseases are enveloped—their virions contain an outer lipid bilayer originating from a host cell. Small molecule antivirals targeting the lipid bilayer cover the broadest spectrum of viruses. In this context, we consider the chemical nature and mechanisms of action of membrane-targeting antivirals. They can affect virions by (1) physically modulating membrane properties to inhibit fusion of the viral envelope with the cell membrane, (2) physically affecting envelope lipids and proteins leading to membrane damage, pore formation and lysis, (3) causing photochemical damage of unsaturated membrane lipids resulting in integrity loss and fusion arrest. Other membrane-active compounds can target host cell membranes involved in virion’s maturation, coating, and egress (endoplasmic reticulum, Golgi apparatus, and outer membrane) affecting these last stages of viral reproduction. Both virion- and host-targeting membrane-active molecules are promising concepts for broad-spectrum antivirals. A panel of approved antivirals would be a superior weapon to respond to and control emerging disease outbreaks caused by new viral strains and variants. Full article

With the advent of Autonomous Vehicles (AV) technology, extensive research around the design of on-demand mobility systems powered by such vehicles is performed. An important part of these studies consists in the evaluation of the economic impact of such systems for involved stakeholders. In this work, a cost–benefit analysis (CBA) is applied to the introduction of AV services in Paris-Saclay, an intercommunity, south of Paris, simulated through MATSim, an agent-based model capable of capturing complex travel behaviors and dynamic traffic interactions. AVs would be implemented as a feeder service, first- and last-mile service to public transit, allowing intermodal trips for travelers. The system is designed to target the challenges of public transport accessibility in periurban areas and high private car use, which the AV feeder service is designed to mitigate. To our knowledge, this study is one of the first CBA analyses of an intermodal AV system relying on an agent-based simulation. The introduction of AV in a periurban environment would generate more pressure on the road network (0.8% to 1.7% increase in VKT for all modes, and significant congestion around train stations) but would improve traveler utilities. The utility gains from the new AV users benefiting from a more comfortable mode offsets the longer travel times from private car users. A Stop-Based routing service generates less congestion than a Door-to-Door routing service, but the access/egress time counterbalances this gain. Finally, in a periurban environment where on-demand AV feeder service would be added to reduce the access and egress cost of public transit, the social impact would be nuanced for travelers (over 99% of gains captured by the 10% of most benefiting agents), but externality would increase. This would benefit some travelers but would also involve additional congestion. In that case, a Stop-Based routing on a constrained network (e.g., existing bus network) significantly improves economic viability and reduces infrastructure costs and would be less impacting than a Door-to-Door service. Full article

As designers push engine efficiency closer to thermodynamic limits, the analysis of flow instabilities developed in a high-pressure turbine (HPT) is crucial to minimizing aerodynamic losses and optimizing secondary air systems. Purge flow, while essential for protecting turbine components from thermal stress, significantly impacts the overall efficiency of the engine and is strictly connected to cavity modes and rim-seal instabilities. This paper presents an experimental investigation of these instabilities in an HPT stage, tested under engine-representative flow conditions in the short-duration turbine rig of the von Karman Institute. As operating conditions significantly influence instability behavior, this study provides valuable insight for future turbine design. Fast-response pressure measurements reveal asynchronous flow instabilities linked to ingress–egress mechanisms, with intensities modulated by the purge rate (PR). The maximum strength is reached at PR = 1.0%, with comparable intensities persisting for higher rates. For lower PRs, the instability diminishes as the cavity becomes unsealed. An analysis based on the cross-power spectral density is applied to quantify the characteristics of the rotating instabilities. The speed of the asynchronous structures exhibits minimal sensitivity to the PR, approximately 65% of the rotor speed. In contrast, the structures’ length scale shows considerable variation, ranging from 11–12 lobes at PR = 1.0% to 14 lobes for PR = 1.74%. The frequency domain analysis reveals a complex modulation of these instabilities and suggests a potential correlation with low-engine-order fluctuations. Full article

This paper presents a detailed analysis of the dynamics of indoor environmental parameters under three simulated fire scenarios in a multi-story building, using the PyroSim platform (based on the Fire Dynamics Simulator—FDS). The study compares two smoke control strategies, organized natural ventilation (a passive system) and mechanical pressurization (an active system), evaluating their influence on temperature, differential pressure, air velocity, heat release rate (HRR), and toxic gas distribution. The simulations revealed that passive systems, relying on the stack effect and vertical natural ventilation, do not ensure the effective control of smoke infiltration into evacuation routes, allowing significant heat accumulation and reduced visibility. The results highlight the superior effectiveness of unidirectional mechanical pressurization in maintaining a stable flow regime, functional visibility, and a safe evacuation environment. A key finding is the transition from static pressure control to velocity-based flow control at the moment of door opening toward the fire source. The results confirm that a dynamically adapted application of mechanical pressurization—synchronized with the opening of access pathways—not only reinforces existing principles for protecting egress routes, but also provides a precise operational approach for optimizing emergency responses in high-rise buildings. Full article

Drug resistance is still one of the main challenges for the treatment of colorectal cancer (CRC). Whilst some resistance mechanisms are well known, from the static therapy success rate, clearly, still much is undiscovered. Intracellular transport mechanisms have attracted attention as having a possible role in drug resistance, and here, the Endosomal Sorting Complex Required for Transport (ESCRT) protein family is studied as a source of drug resistance modulation using human CRC cell lines and clinical material. From an initial screening of ESCRT proteins in a panel of 10 CRC wild-type cell lines using immunoblotting, Vacuolar Protein Sorting-Associated Protein A4 (VPS4A) was identified as being consistently highly expressed, and it was selected for further investigation. Immunohistopathological evaluation in a small panel of CRC patient samples demonstrated high expression in the tumor epithelium compared to normal intestinal epithelium. The knockdown of VPS4A resulted in enhanced sensitivity of cells to oxaliplatin, and it was subsequently seen that oxaliplatin-resistant sublines had significantly higher VPS4A expression than their wild-type variants. In addition, it was demonstrated that a small molecule inhibitor of VPS4A, aloperine, could interact synergistically with oxaliplatin to enhance its sensitivity in an oxaliplatin-resistant cell line. We hypothesize from initial RNA sequencing analysis that the mechanism of action of VPS4A modulation is through depleting levels of the drug efflux transporter MRP2 in the cell, preventing oxaliplatin egress and increasing cell exposure to the drug. The evidence presented here thus indicates that ESCRT machinery, specifically VPS4A, may act as a modulator of oxaliplatin resistance in CRC. Full article

Polyomaviruses have the potential to cause significant morbidity not only in transplant medicine, but also in other forms of disease or variants of immunosuppression. In kidney transplant recipients or recipients of human stem cell transplants, the BK-Virus is the major proponent of manifestations such as BKPyV-associated nephropathy or hemorrhagic cystitis. As no polyomavirus-specific drug with proven in vivo effects has been developed so far, methods to screen for such drugs are important. This work describes the establishment of a virus-secreting cell line. By infecting a pre-established monkey kidney cell line (COS-1) with a non-rearranged human BK polyomavirus isolated from a kidney transplant patient suffering from BKPyV-associated nephropathy, a continuously replicating cell type with consistent virus secretion could be established and was termed COSSA. Measurements of BKPyV replication, virion production, and secretion were performed both intracellularly and in the cell supernatant. Viral proteins such as VP1 and LTAg were accurately tracked by confocal microscopy, as well as by immunoblot and qPCR. An intracellular flow cytometry (FACS) assay detecting VP1 protein was established and revealed an expanded range of positive intracellular signals. The viruses produced proved to be infectious in human tubular epithelial cell lines. Long-range sequencing of the COSSA genome using Oxford Nanopore Technology revealed a total of five distinct BKPyV integration events. One integration of a partial BKPyV genome was located upstream of the epidermal growth factor receptor gene. The second and third, both truncated forms of integration, were close to histocompatibility gene locuses, while the fourth was characterized by a ninefold and the fifth by a fourfold tandem repeat of the BKPyV genome. From both of the repeat forms, virus replicates were derived showing deletions/duplications on early and late genes and inversions within the non-coding control region (NCCR). This pattern of repetitive viral genome integration is a potential key driver of enhanced viral replication and increased virion assembly, ultimately supporting efficient virus egress. Quantitative PCR analysis confirmed the release of approximately 10⁸/mL viral units per 48 h from 2 × 10⁵ COSSA cells into the culture supernatant. Notably, the NCCR region of the most frequent copies of circular virus and the integrated tetrameric tandem repeat exhibited a rearranged configuration, which may contribute to the observed high replication dynamics. The establishment of a consistent methodology to generate and secrete BKPyV from a cell line is expected to significantly facilitate antiviral drug development. Full article

Innovative demand-responsive transport services are spreading in most urban areas, allowing dynamic matching between demand and supply and enabling travellers to request shared rides in real-time via mobile applications. They are used both as an alternative to public transport and as an access/egress leg to mass transit stations, i.e., acting as a feeder service. In low-demand areas and small-sized cities, it is often difficult to provide effective and cost-efficient public transport, thus resulting in an extensive use of private vehicles. Using an agent-based modelling approach, this study compares the performance of fixed-route transit (FRT) and demand-responsive transit (DRT), where optional stops can be activated on demand. The aim is to identify the conditions allowing DRT to become more advantageous than FRT in small-sized cities, both for travellers and the transport operator. A real-time matching algorithm identifies optimal trip chains (i.e., public transport lines; pick-up, drop-off and transfer stops; and time windows) for travel requests, dynamically updating vehicles’ routes and schedules. The model is applied to the city of Caltanissetta, Italy, where a transit service with six FRT urban lines is currently operating. Travel patterns were reconstructed from thousands of travel requests collected by a Mobility-as-a-Service platform within one-year. The main findings demonstrate the benefits of DRT in providing a higher quality of service, reducing riding times for passengers, and enhancing service efficiency without burdening operating costs. The DRT reduced the vehicle-kilometres travelled by up to 5% compared to FRT while decreasing passenger ride times by approximately 10%. An economic analysis showed reductions in operator unit costs of up to 3.4% for low-demand rates, confirming the advantages of flexible operations in small-sized cities. Full article

Hendra virus (HeV) is a highly pathogenic member of the Henipavirus genus (family Paramyxoviridae, order Mononegavirales), for which all basic replication processes are located in the cytoplasm. The HeV matrix (M) protein plays essential roles in viral assembly and budding at the plasma membrane, but also undergoes dynamic nuclear and nucleolar trafficking, accumulating in nucleoli early in infection, before relocalising to the plasma membrane. We previously showed that M targets sub-nucleolar compartments—the fibrillar centre (FC) and dense fibrillar component (DFC)—to modulate rRNA biogenesis by mimicking a process occurring during a nucleolar DNA-damage response (DDR). Here, we show that M protein sub-nucleolar localisation is regulated by ubiquitination, which controls its redistribution between the FC-DFC and granular component (GC). The mutagenesis of a conserved lysine (K258) reported to undergo ubiquitination, combined with the pharmacological modulation of ubiquitination, indicated that a positive charge at K258 is required for M localisation to the FC-DFC, while ubiquitination regulates subsequent egress from the FC-DFC to the GC. M proteins from multiple Henipaviruses exhibited similar ubiquitin-dependent sub-nucleolar trafficking, indicating a conserved mechanism. These findings reveal a novel mechanism regulating viral protein transport between phase-separated sub-nucleolar compartments and highlight ubiquitination as a key modulator of intra-nucleolar trafficking. Full article

Sphingosine 1 phosphate receptor (S1PR) modulators are the latest drug class to have received approval for the treatment of ulcerative colitis, and have brought a new mechanism of action to this landscape. They target immune cell trafficking, specifically the egress of lymphocytes from lymph nodes to the bloodstream, and have proven to be an efficacious and safe anti-inflammatory mechanism. This narrative review aims to distil the key trial data on the efficacy and safety of ozanimod and etrasimod, the two S1PR modulators currently licensed for use in UC. We discuss the higher response rates in the advanced therapy naive versus exposed subgroups. We summarise their safety profiles, taking into consideration open label extension data. Finally, we consider where this class of drugs may be best placed in the treatment landscape and also provide a practical guide for their use in clinical practice. Full article

In this paper, a panel smooth transition regression model is used to examine the nonlinear effects of financial globalization on energy security. These effects are examined in 123 countries for the period of 2000–2018. Control variables are armed forces, industrialization rate, trade value share, and urbanization rate, and the conversion variable is the financial globalization index in the following year. The results of the financial globalization effects can be obtained from both time and space. The results show that financial globalization has a positive nonlinear effect on energy security. When the logarithm of financial globalization in the previous year exceeds 0.0467, the coefficient between financial globalization and energy security will decrease from 0.0467 to 0.0209. Temporal variation analyses show that the positive effect followed a “decrease, increase, decrease” trend between 2000 and 2018. Spatial variation analyses show that the positive effect is greatest in Oceania and the Americas (with an effect coefficient of 0.0467) and smallest in Europe (with an effect coefficient of 0.0391). According to the results of the regional heterogeneity research, the Organization of the Petroleum Exporting Countries (OPEC) countries see a stronger nonlinear impact of financial globalization on energy security than non-OPEC countries. Full article

This study introduces a new metro train demand model that simultaneously captures both aggregate ridership from automated fare collection (AFC) data and disaggregate consideration propensities, using individual survey data from Chennai, India. This joint framework produces more accurate aggregate demand estimates than traditional OLS (R² improves from 0.67 to 0.75), as it is able to capture the complex and non-linear relationship between disaggregate consideration probability, reflecting potential demand, and aggregate footfall, reflecting realized demand. It is observed that increasing the consideration probability enhances the footfall overall. However, some locations exhibit an opposing trend between consideration and footfall (low consideration but high footfall, or vice versa). Also, the sets of influential factors vary across these two dimensions. For instance, individual-level variables (income and out-of-vehicle travel time) and multi-modal connectivity features (presence of an airport and multimodal hubs near the metro) play a key role in footfall. In contrast, consideration probability is primarily influenced by access time, cost, and egress distance. Furthermore, factors influencing consideration probability (walkability, train service quality, and first–last–mile connectivity) vary across segments (based on vehicle unavailability, exclusive vehicle availability, and limited vehicle availability). Evidence of selection bias among metro riders, non-normality, and intra-person variability effects in footfall is observed. From a policy perspective, neglecting the disaggregate consideration effects on realized aggregate demand, i.e., footfall models, can overestimate the role of metro costs and out-of-vehicle travel time. In addition, the ridership levels of the metro are overestimated at higher metro fare levels. The new model illustrates that applying location-specific and dimension-specific policy interventions can be more effective than uniform area-wide policies for enhancing the user base and realized ridership. Full article

Toxoplasma gondii is a protozoan parasite capable of establishing chronic infections, with potential reactivation in immunocompromised individuals. However, the molecular mechanisms governing tachyzoite-to-bradyzoite differentiation remain incompletely understood. Previous studies have identified AP2 transcription factors as key regulators of this developmental switch. In this study, we investigated the role of the AP2 factor AP2X-8. Immunofluorescence analysis revealed that AP2X-8 is constitutively expressed in the nucleus of both tachyzoite and bradyzoite stages. Using CRISPR-Cas9-mediated homologous recombination, we successfully generated an ap2X-8 knockout strain. Phenotypic assays including plaque formation, invasion, replication, and egress, and bradyzoite differentiation assays, were then performed to assess the impact of ap2X-8 deletion. Our analyses showed that the loss of ap2X-8 significantly impaired plaque formation and intracellular replication, while invasion and egress were unaffected. Furthermore, ap2X-8 knockout enhanced bradyzoite differentiation in vitro. Despite these changes, deletion of ap2X-8 did not alter parasite virulence in a mouse infection model. These findings demonstrate that AP2X-8 is an important regulator of T. gondii tachyzoite growth and bradyzoite differentiation, offering new insights into the parasite’s developmental regulation. Full article

Hypercapnia, the elevation of CO₂ in blood and tissue, is a risk factor for mortality in patients with severe lung disease and pulmonary infections. We previously showed that hypercapnia increases viral replication and mortality in mice infected with influenza A virus (IAV). Elevated CO₂ also augmented cholesterol content and pseudo-SARS-CoV-2 entry in bronchial epithelial cells. Interestingly, cellular cholesterol facilitates IAV uptake, replication, assembly, and egress from cells. Here, we report that hypercapnia increases viral protein expression in airway epithelium of mice infected with IAV. Elevated CO₂ also enhanced IAV adhesion and internalization, viral protein expression, and viral replication in bronchial epithelial cells. Hypercapnia increased the expression and activation of the transcription factor sterol-regulatory element binding protein 2 (SREBP2), resulting in elevated expression of cholesterol synthesis enzymes, decreased expression of a cholesterol efflux transporter, and augmented cellular cholesterol. Moreover, reducing cellular cholesterol with an SREBP2 inhibitor or statins blocked hypercapnia-induced increases in viral adhesion and internalization, viral protein expression, and IAV replication. Inhibitors of mTOR and Akt also blocked the effect of hypercapnia on viral growth. Our findings suggest that targeting cholesterol synthesis and/or mTOR/Akt signaling may hold promise for reducing susceptibility to influenza infection in patients with advanced lung disease and hypercapnia. Full article

This study explores the impact of key evacuation features on occupant safety in complex buildings with underground connections in Seoul, the city with the highest concentration of such buildings in the country. By analyzing factors like exit spacing, exit width, stairwell distances, and stairway configurations, the study assesses evacuation safety using fire and evacuation simulations, comparing available safe egress time (ASET) with required safe egress time (RSET). Reducing interior exit facility spacing from the legal standard of 100 m to 50 m improved evacuation time by 77.5% (from 36 min to 8 min and 7 s), with a further reduction to 40 m improving performance by an additional 23.3% (to 6 min and 13 s). In downward evacuations, reducing the walking distance to exterior exits from over 50 m to 30 m cut evacuation time by at least 59.9% (from 23 min and 55 s to 9 min and 35 s), ensuring successful evacuations. These findings demonstrate that optimizing evacuation routes, addressing bottlenecks, and improving evacuation feature standards can significantly enhance safety and minimize casualties. By adjusting building design and fire safety regulations, these optimizations promote resilient urban infrastructure, reduce disaster-related socio-economic impacts, and inform evidence-based policies, offering valuable insights for policymakers and guiding future improvements in fire safety and evacuation protocols. Full article

The open reading frame 103 (p48) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) is one of the 38 core baculovirus genes. p48 has been shown to be essential for the production of infectious budded virions (BVs), nuclear egress of nucleocapsids, envelopment of the nucleocapsid, and embedding of occlusion-derived virions (ODVs) into occlusion bodies (OBs). However, the structure–function relationship of P48 remains unclear. In this study, we showed that four conserved cysteines (C127, C130, C138, and C141) in P48 may form a zinc finger motif based on a predicted structure analysis, and we investigated the roles of these cysteines in P48 function. AcMNPV bacmids lacking p48 or containing mutated p48 were generated. Transfection/infection assays showed that C127, C130, C138, and C141 in P48 were crucial for infectious BV production. Electron microscopy analysis further confirmed that these four cysteines played critical roles in the transport of nucleocapsids out of the nucleus for BV production, and in ODV envelopment. These results demonstrate that the conserved cysteines C127, C130, C138, and C141, related to the putative zinc finger motif, are critical for P48 function in baculovirus infection. Full article