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The airway surface liquid (ASL) plays a crucial role in lung defense mechanisms, and its composition and volume are regulated by the airway epithelium. The cystic fibrosis transmembrane conductance regulator (CFTR) is abundantly expressed in a rare airway epithelial cell type called an ionocyte. Recently, we demonstrated that ionocytes can increase liquid absorption through apical CFTR and basolateral barttin/chloride channels, while airway secretory cells mediate liquid secretion through apical CFTR channels and basolateral NKCC1 transporters. Th2-driven (IL-4/IL-13) airway diseases, such as asthma, cause goblet cell metaplasia, accompanied by increased mucus production and airway secretions. In this study, we investigate the effect of IL-13 on chloride and liquid transport performed by ionocytes. IL-13 treatment of human airway epithelia was associated with reduced epithelial liquid absorption rates and increased ASL volume. Additionally, IL-13 treatment reduced the abundance of CFTR-positive ionocytes and increased the abundance of CFTR-positive secretory cells. Increasing ionocyte abundance attenuated liquid secretion caused by IL-13. Finally, CFTR-positive ionocytes were less common in asthma and COPD and associated with airflow obstruction. Our findings suggest that loss of CFTR in ionocytes contributes to the liquid secretion observed in IL-13-mediated airway diseases.

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Winter cover crop performance metrics (i.e., vegetative biomass quantity and quality) affect ecosystem services provisions, but they vary widely due to differences in agronomic practices, soil properties, and climate. Cereal rye (Secale cereale) is the most common winter cover crop in the United States due to its winter hardiness, low seed cost, and high biomass production. We compiled data on cereal rye winter cover crop performance metrics, agronomic practices, and soil properties across the eastern half of the United States. The dataset includes a total of 5,695 cereal rye biomass observations across 208 site-years between 2001-2022 and encompasses a wide range of agronomic, soils, and climate conditions. Cereal rye biomass values had a mean of 3,428 kg ha-1, a median of 2,458 kg ha-1, and a standard deviation of 3,163 kg ha-1. The data can be used for empirical analyses, to calibrate, validate, and evaluate process-based models, and to develop decision support tools for management and policy decisions.

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Introduction: The frequent use of pesticides is currently considered a cause of environmental pollution due to the high rate of entry of these substances into agroecosystems. This constitutes a risk for the species that inhabit these ecosystems, in particular anurans whose characteristics make them prone to exposure to and interaction with environmental pollutants. Objective: To report the occurrence of abnormalities in larvae of the common toad Rhinella arenarum inhabiting ponds surrounded by agroecosystems. Methods: In two consecutive springs (2015 and 2016), reproductive events of common toads were monitored in temporary pond systems in agricultural and non-agricultural areas, located near the city of La Plata (Buenos Aires, Argentina). The physicochemical parameters of the ponds were measured, and the stage of each reproductive event was recorded, such as the numbers of adult toads, amplexus and clutches. In the laboratory, the larvae were measured and photographed, their stage of development was recorded, and their morphology was examined under a stereomicroscope. Representative samples (normal and abnormal) from each pond studied were processed for histopathological analysis. Results: In the field studies carried out on a population of R. arenarum collected in an agroecosystem, a lower number of reproductive adults and clutches were observed in relation to the population of a non-agricultural pond. A total of 1 910 larvae were collected: 529 and 1 381 larvae from ponds located in non-agricultural and agricultural areas, respectively. Larvae from the agroecosystem showed two types of abnormalities: severe tail flexure and abdominal bloating. In addition, five degrees of severity could be determined in relation to abdominal bloating. Conclusions: This work reports the high frequency and severity of abnormalities observed in the early stages of R. arenarum larvae living within an agroecosystem, providing evidence of the negative impact that agricultural activities cause on aquatic ecosystems surrounded by farming areas.

Introducción: El uso frecuente de plaguicidas es considerado actualmente una causa de contaminación ambiental debido a las altas tasas de ingreso de estas sustancias a los agroecosistemas. Esta situación es un riesgo para las especies que habitan en estos ecosistemas, en particular los anuros cuyas características los hacen propensos a la exposición e interacción con contaminantes ambientales. Objetivo: Informar la presencia de anormalidades en larvas del sapo común Rhinella arenarum que habitan en estanques rodeados por un agroecosistema. Métodos: En dos primaveras consecutivas (2015 y 2016), se monitorearon los eventos reproductivos del sapo común proveniente de sistemas de estanques temporales ubicados en zonas agrícolas y no agrícolas, cerca de la ciudad de La Plata (Buenos Aires, Argentina). Se midieron los parámetros fisicoquímicos de los estanques y se registraron las etapas de cada evento reproductivo como el número de sapos adultos, amplexos y nidadas. En el laboratorio, las larvas fueron medidas y fotografiadas, se registró su estado de desarrollo y se examinó la morfología de cada una bajo microscopio estereoscópico. Se procesaron muestras representativas (normales y anormales) de cada estanque estudiado para análisis histopatológico. Resultados: En la población de R. arenarum que vive dentro de un agroecosistema, se observó un menor número de adultos reproductores y puestas en relación con la del estanque en la zona no agrícola. Se recolectaron un total de 1 910 larvas: 529 y 1 381 larvas de estanques ubicados en zonas no agrícolas y agrícolas, respectivamente. Las larvas del agroecosistema mostraron dos tipos de anormalidades: severa flexión de la cola y distensión abdominal. Además, se pudo determinar cinco grados de gravedad en relación con la distensión abdominal. Conclusiones: Una alta frecuencia y severidad de anormalidades en los estadios tempranos de larvas de R. arenarum que viven dentro de un agroecosistema proporciona evidencia del impacto negativo que las actividades agrícolas causan en los ecosistemas acuáticos rodeados por áreas de cultivo.

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The volume and composition of a thin layer of liquid covering the airway surface defend the lung from inhaled pathogens and debris. Airway epithelia secrete Cl- into the airway surface liquid through cystic fibrosis transmembrane conductance regulator (CFTR) channels, thereby increasing the volume of airway surface liquid. The discovery that pulmonary ionocytes contain high levels of CFTR led us to predict that ionocytes drive secretion. However, we found the opposite. Elevating ionocyte abundance increased liquid absorption, whereas reducing ionocyte abundance increased secretion. In contrast to other airway epithelial cells, ionocytes contained barttin/Cl- channels in their basolateral membrane. Disrupting barttin/Cl- channel function impaired liquid absorption, and overexpressing barttin/Cl- channels increased absorption. Together, apical CFTR and basolateral barttin/Cl- channels provide an electrically conductive pathway for Cl- flow through ionocytes, and the transepithelial voltage generated by apical Na+ channels drives absorption. These findings indicate that ionocytes mediate liquid absorption, and secretory cells mediate liquid secretion. Segregating these counteracting activities to distinct cell types enables epithelia to precisely control the airway surface. Moreover, the divergent role of CFTR in ionocytes and secretory cells suggests that cystic fibrosis disrupts both liquid secretion and absorption.

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Genomic selection (GS) is transforming plant and animal breeding, but its practical implementation for complex traits and multi-environmental trials remains challenging. To address this issue, this study investigates the integration of environmental information with genotypic information in GS. The study proposes the use of two feature selection methods (Pearson's correlation and Boruta) for the integration of environmental information. Results indicate that the simple incorporation of environmental covariates may increase or decrease prediction accuracy depending on the case. However, optimal incorporation of environmental covariates using feature selection significantly improves prediction accuracy in four out of six datasets between 14.25% and 218.71% under a leave one environment out cross validation scenario in terms of Normalized Root Mean Squared Error, but not relevant gain was observed in terms of Pearson´s correlation. In two datasets where environmental covariates are unrelated to the response variable, feature selection is unable to enhance prediction accuracy. Therefore, the study provides empirical evidence supporting the use of feature selection to improve the prediction power of GS.

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While sparse testing methods have been proposed by researchers to improve the efficiency of genomic selection (GS) in breeding programs, there are several factors that can hinder this. In this research, we evaluated four methods (M1-M4) for sparse testing allocation of lines to environments under multi-environmental trails for genomic prediction of unobserved lines. The sparse testing methods described in this study are applied in a two-stage analysis to build the genomic training and testing sets in a strategy that allows each location or environment to evaluate only a subset of all genotypes rather than all of them. To ensure a valid implementation, the sparse testing methods presented here require BLUEs (or BLUPs) of the lines to be computed at the first stage using an appropriate experimental design and statistical analyses in each location (or environment). The evaluation of the four cultivar allocation methods to environments of the second stage was done with four data sets (two large and two small) under a multi-trait and uni-trait framework. We found that the multi-trait model produced better genomic prediction (GP) accuracy than the uni-trait model and that methods M3 and M4 were slightly better than methods M1 and M2 for the allocation of lines to environments. Some of the most important findings, however, were that even under a scenario where we used a training-testing relation of 15-85%, the prediction accuracy of the four methods barely decreased. This indicates that genomic sparse testing methods for data sets under these scenarios can save considerable operational and financial resources with only a small loss in precision, which can be shown in our cost-benefit analysis.

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Emulsions are critical across a broad spectrum of industries. Unfortunately, emulsification requires a significant driving force for droplet dispersion. Here, we demonstrate a mechanism of spontaneous droplet formation (emulsification), where the interfacial solute flux promotes droplet formation at the liquid-liquid interface when a phase transfer agent is present. We have termed this phenomenon fluxification. For example, when HAuCl4 is dissolved in an aqueous phase and [NBu4][ClO4] is dissolved in an oil phase, emulsion droplets (both water-in-oil and oil-in-water) can be observed at the interface for various oil phases (1,2-dichloroethane, dichloromethane, chloroform, and nitrobenzene). Emulsification occurs when AuCl4- interacts with NBu4+, a well-known phase-transfer agent, and transfers into the oil phase while ClO4- transfers into the aqueous phase to maintain electroneutrality. The phase transfer of SCN- and Fe(CN)63- also produce droplets. We propose a microscopic mechanism of droplet formation and discuss design principles by tuning experimental parameters.

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Nanodroplet-mediated electrodeposition is a reliable method for electrodepositing nanoparticles by confining a small amount of metal-salt precursor in water nanodroplets (radius ∼400 nm) suspended in an oil continuous phase. This technique provides a great advantage in terms of nanoparticle size, morphology, and porosity. For an electrochemical reaction to proceed in the aqueous nanodroplet, the electroneutrality condition must be maintained. Classically, [NB4][ClO4] or a comparable salt is added to the oil continuous phase to maintain charge balance. Unfortunately, the presence of this salt in the oil phase causes some metal salts, such as HAuCl4, to phase transfer, disallowing the formation of gold nanoparticles. Here, we demonstrate the partitioning of HAuCl4 is orders of magnitude lower using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) when LiClO4 is added to the nanodroplet phase and [NBu4][ClO4] is not added to the continuous phase. This simple change allows for the electrodeposition of gold nanoparticles. Scanning electron microscopy shows the morphology and size distribution of gold nanoparticles obtained at different concentrations of LiClO4. Transmission electron microscopy in selected diffraction mode was used and it determined the gold nanoparticles obtained are polycrystalline with miller indices of (222) and (200). This work widens the variety of nanoparticles that can be electrodeposited from nanodroplets for applications in energy storage and conversion, photoelectrochemistry, and biosensing.

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Maintaining the integrity of the cell plasma membrane (PM) is critical for the survival of cells. While an efficient PM repair machinery can aid survival of healthy cells by preventing influx of extracellular calcium, it can also constitute an obstacle in drug delivery and photothermal therapy. We show how nanoscopic holes can be created in a controlled fashion to the cell's plasma membrane, thus allowing identification of molecular components which have a pivotal role in PM repair. Cells are punctured by laser induced local heating of gold nanostructures at the cell surface which causes nano-ruptures in cellular PMs. Recruitment of annexin V near the hole is found to locally reshape the ruptured plasma membrane. Experiments using model membranes, containing recombinant annexin V, provide further biophysical insight into the ability of annexin V to reshape edges surrounding a membrane hole. The thermoplasmonic method provides a general strategy to monitor the response to nanoscopic injuries to the cell surface which offer new insight into how cells respond to photothermal treatment.

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Lack of CFTR (cystic fibrosis transmembrane conductance regulator) affects the transcriptome, composition, and function of large and small airway epithelia in people with advanced cystic fibrosis (CF); however, whether lack of CFTR causes cell-intrinsic abnormalities present at birth versus inflammation-dependent abnormalities is unclear. We performed a single-cell RNA-sequencing census of microdissected small airways from newborn CF pigs, which recapitulate CF host defense defects and pathology over time. Lack of CFTR minimally affected the transcriptome of large and small airways at birth, suggesting that infection and inflammation drive transcriptomic abnormalities in advanced CF. Importantly, common small airway epithelial cell types expressed a markedly different transcriptome than corresponding large airway cell types. Quantitative immunohistochemistry and electrophysiology of small airway epithelia demonstrated basal cells that reach the apical surface and a water and ion transport advantage. This single cell atlas highlights the archetypal nature of airway epithelial cells with location-dependent gene expression and function.

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OBJECTIVE: To analyze the caesarean deliveries attended in our hospital, grouping them according to the Robson Classification System and to establish measures in order to reduce caesarean delivery rates. METHOD: Prospective study of all the deliveries attended at Hospital Doctor Peset in 2019 using the Robson classification. RESULTS: A total of 1113 births have been analyzed with a total cesarean section rate of 25.3%. The largest contribution to the total cesarean delivery rate with 34.4% was group 2A (nulliparous women with a single fetus in cephalic presentation, 37 weeks or more pregnant who started labor by induction). Secondly, group 5 (multiparous women with at least previous cesarean section, with single cephalic fetus, 37 weeks or more pregnant) which represents the 20.1% of the total. Inductions in nulliparas multiply the cesarean section rate by 3 compared to nulliparas that initiate labor spontaneously. CONCLUSIONS: Robsons classification is a tool that allows to easily classify and analyze the groups in which to implement measures to reduce the number of caesarean sections performed. Analyzing the induction indications and reviewing action protocols could suppose a substantial decrease in the caesarean section rate in our center.

OBJETIVO: Analizar las cesáreas realizadas en nuestro centro agrupándolas según la clasificación de Robson para establecer medidas que permitan reducir la tasa de cesáreas. MÉTODO: Auditoría prospectiva de los nacimientos asistidos en el Hospital Doctor Peset en el año 2019 mediante la clasificación de Robson. RESULTADOS: Se han analizado 1113 nacimientos con una tasa de cesárea del 25.3%. El grupo que más contribuyó al total de cesáreas realizadas, con un 34.4%, fue el 2A (nulíparas con feto único en presentación cefálica, de 37 semanas o más de embarazo, que iniciaron el parto mediante inducción). En segundo lugar, el grupo 5 (multíparas con al menos una cesárea previa, con un feto único en presentación cefálica, de 37 semanas o más de embarazo), con un 20.1%. Las inducciones en nulíparas multiplican por tres la tasa de cesárea respecto a las nulíparas que inician el trabajo de parto de manera espontánea. CONCLUSIONES: La clasificación de Robson es una herramienta que permite clasificar y analizar de manera sencilla los grupos en los que implantar medidas para reducir el número de cesáreas realizadas. Analizar las indicaciones de inducción y revisar los protocolos de actuación podría suponer una disminución sustancial en la tasa de cesáreas en nuestro centro.

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Large-scale ecotoxicological studies have technical and ethical limitations, both related to the need to expose large numbers of individuals to potentially harmful compounds. The computational modeling is a complementary useful and predictive tool that overcomes these limitations. Considering the increasing interest in the effects of pesticides on behavioral traits, the aim of this study was to evaluate the effects of chlorpyrifos (CPF) on intra- and inter-specific interactions of anuran tadpoles, complementing traditional ecotoxicological tools with a theoretical analysis verified by computational simulations. Experiments were developed under two consecutive phases: a first phase of exposure (treated and control group), and a second phase of interactions. The second phase consisted of evaluating the effects of CPF on intra- and inter-specific interactions of exposed C. ornata (Co) tadpoles acting as predators and unexposed Rhinella fernandezae (Rf) tadpoles acting as prey (Experiment I), under different predator-prey proportions (0/10 = 0Co-10Rf, 2/8, 4/6, 6/4, 8/2, 10/0). Also, intraspecific interactions of three Co tadpoles under different conditions of exposure were evaluated (Experiment II: 3 exposed Co, 2 exposed Co/1 non-exposed, 1 exposed Co/2 non-exposed). During the exposure phase, chlorpyrifos induced significant mortality from 48 h (48 h: p < 0.05, 72 h-96 h: p < 0.001), irregular swimming, tail flexure, and the presence of subcutaneous air. Also, it induced effects on the sounds emitted after 96 h of exposure, registering a smaller number of pulses and higher dominant frequencies, and altered intra- and inter-specific interactions. During the interaction phase, the larvae continued to show sound effects, however, the antipredator mechanism continued to be operating and efficient. Finally, it was possible to model the behavior of the larvae under the effects of chlorpyrifos. We conclude that experimental data and computational modeling matched. Therefore, computational simulation is a valuable ecotoxicological tool that provides new information and allows prediction of natural processes.

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Bronchogenic cysts are congenital anomalies of the respiratory tract, which can affect children or adults. In adults, these cysts may be asymptomatic or present with symptoms, most often pain or cough. Intrapericardial cysts are rare and represent 27% of bronchogenic cysts. We present the case of a symptomatic adult with an intrapericardial bronchogenic cyst, who underwent surgical resection of the cyst, which successfully relieved his symptoms.

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Submucosal glands (SMGs) are a prominent structure that lines human cartilaginous airways. Although it has been assumed that SMGs contribute to respiratory defense, that hypothesis has gone without a direct test. Therefore, we studied pigs, which have lungs like humans, and disrupted the gene for ectodysplasin (EDA-KO), which initiates SMG development. EDA-KO pigs lacked SMGs throughout the airways. Their airway surface liquid had a reduced ability to kill bacteria, consistent with SMG production of antimicrobials. In wild-type pigs, SMGs secrete mucus that emerges onto the airway surface as strands. Lack of SMGs and mucus strands disrupted mucociliary transport in EDA-KO pigs. Consequently, EDA-KO pigs failed to eradicate a bacterial challenge in lung regions normally populated by SMGs. These in vivo and ex vivo results indicate that SMGs are required for normal antimicrobial activity and mucociliary transport, two key host defenses that protect the lung.

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Lentiviral-mediated integration of a CFTR transgene cassette into airway basal cells is a strategy being considered for cystic fibrosis (CF) cell-based therapies. However, CFTR expression is highly regulated in differentiated airway cell types and a subset of intermediate basal cells destined to differentiate. Since basal stem cells typically do not express CFTR, suppressing the CFTR expression from the lentiviral vector in airway basal cells may be beneficial for maintaining their proliferative capacity and multipotency. We identified miR-106b as highly expressed in proliferating airway basal cells and extinguished in differentiated columnar cells. Herein, we developed lentiviral vectors with the miR-106b-target sequence (miRT) to both study miR-106b regulation during basal cell differentiation and detarget CFTR expression in basal cells. Given that miR-106b is expressed in the 293T cells used for viral production, obstacles of viral genome integrity and titers were overcome by creating a 293T-B2 cell line that inducibly expresses the RNAi suppressor B2 protein from flock house virus. While miR-106b vectors effectively detargeted reporter gene expression in proliferating basal cells and following differentiation in the air-liquid interface and organoid cultures, the CFTR-miRT vector produced significantly less CFTR-mediated current than the non-miR-targeted CFTR vector following transduction and differentiation of CF basal cells. These findings suggest that miR-106b is expressed in certain airway cell types that contribute to the majority of CFTR anion transport in airway epithelium.

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The pH of airway surface liquid (ASL) is a key factor that determines respiratory host defense; ASL acidification impairs and alkalinization enhances key defense mechanisms. Under healthy conditions, airway epithelia secrete base ([Formula: see text]) and acid (H+) to control ASL pH (pHASL). Neutrophil-predominant inflammation is a hallmark of several airway diseases, and TNFα and IL-17 are key drivers. However, how these cytokines perturb pHASL regulation is uncertain. In primary cultures of differentiated human airway epithelia, TNFα decreased and IL-17 did not change pHASL. However, the combination (TNFα+IL-17) markedly increased pHASL by increasing [Formula: see text] secretion. TNFα+IL-17 increased expression and function of two apical [Formula: see text] transporters, CFTR anion channels and pendrin Cl-/[Formula: see text] exchangers. Both were required for maximal alkalinization. TNFα+IL-17 induced pendrin expression primarily in secretory cells where it was coexpressed with CFTR. Interestingly, significant pendrin expression was not detected in CFTR-rich ionocytes. These results indicate that TNFα+IL-17 stimulate [Formula: see text] secretion via CFTR and pendrin to alkalinize ASL, which may represent an important defense mechanism in inflamed airways.

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The effects of climate change together with the projected future demand represents a huge challenge for wheat production systems worldwide. Wheat breeding can contribute to global food security through the creation of genotypes exhibiting stress tolerance and higher yield potential. The objectives of our study were to (i) estimate the annual grain yield (GY) genetic gain of High Rainfall Wheat Yield Trials (HRWYT) grown from 2007 (15th HRWYT) to 2016 (24th HRWYT) across international environments, and (ii) determine the changes in physiological traits associated with GY genetic improvement. The GY genetic gains were estimated as genetic progress per se (GYP) and in terms of local checks (GYLC). In total, 239 international locations were classified into two groups: high- and low-rainfall environments based on climate variables and trial management practices. In the high-rainfall environment, the annual genetic gains for GYP and GYLC were 3.8 and 1.17 % (160 and 65.1 kg ha-1 yr-1), respectively. In the low-rainfall environment, the genetic gains were 0.93 and 0.73 % (40 and 33.1 kg ha-1 yr-1), for GYP and GYLC respectively. The GY of the lines included in each nursery showed a significant phenotypic correlation between high- and low-rainfall environments in all the examined years and several of the five best performing lines were common in both environments. The GY progress was mainly associated with increased grain weight (R2 = 0.35 p < 0.001), days to maturity (R2 = 0.20, p < 0.001) and grain filling period (R2 = 0.06, p < 0.05). These results indicate continuous GY genetic progress and yield stability in the HRWYT germplasm developed and distributed by CIMMYT.

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Several DNA viruses have evolved antagonists to inhibit the cyclic GMP-AMP synthase (cGAS)-stimulator of interferon genes (STING) DNA-sensing immune pathway. This includes DNA viral oncogenes that antagonize the cGAS-STING pathway by binding STING through the LxCxE motif. The 293T human cells are widely used in biology studies as they are highly transfectable. While parental 293 cells express high levels of STING, 293T cells lack STING and are unable to induce interferon antiviral responses to cytosolic DNA. Additionally, 293T cells express the SV40 polyomavirus large T antigen (LT) which enhances the replication of transfected DNA plasmids carrying the SV40 origin of replication. Since SV40 LT also encodes the LxCxE motif, the lack of STING expression in 293T cells is commonly assumed to be due to SV40 large T antigen. We find that SV40 LT does not alter exogenously expressed and endogenous levels of STING protein. We show that STING transcription is suppressed in 293T cells but is not driven by SV40. This study also revealed that SV40 LT does indeed inhibit cGAS-STING interferon induction, but through a mechanism distinct from other DNA virus oncogenes. Collectively, these results indicate that while SV40 LT can inhibit cGAS-STING interferon induction, it does so in an unanticipated manner.

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