The Foot-and-Mouth Disease Virus Lb Protease Cleaves Intracellular Transcription Factors STAT1 and STAT2 to Antagonize IFN-ß-Induced Signaling.

Foot-and-mouth disease virus (FMDV) is the causative agent of foot-and-mouth disease, one of the most highly infectious animal viruses throughout the world. The JAK-STAT signaling pathway is a highly conserved pathway for IFN-ß-induced antiviral gene expression. Previous studies have shown that FMDV can strongly suppress the innate immune response. Moreover, although STAT1 and STAT2 (STAT1/2) have been well established in JAK-STAT signaling-induced antiviral gene expression, whether FMDV proteins inhibit IFN-ß-induced JAK-STAT signaling remains poorly understood. In this study, we described the Lb leader protease (Lbpro) of FMDV as a candidate for inhibiting IFN-ß-induced signaling transduction via directly interacting with STAT1/2. We further showed that Lbpro colocalized with STAT1/2 to inhibit their nuclear translocation. Importantly, Lbpro cleaved STAT1/2 to inhibit IFN-ß-induced signal transduction, whereas the catalytically inactive mutant of LC51A (Lbpro with cysteine substituted with alanine at amino acid residue 51) had no effect on the stability of STAT1/2 proteins. The cleavage of the STAT1/2 proteins was also determined during FMDV infection in vitro. Lbpro could cleave the residues between 252 and 502 aa for STAT1 and the site spanning residues 140 - 150 aa (QQHEIESRIL) for STAT2. The in vivo results showed that Lbpro can cleave STAT1/2 in pigs. Overall, our findings suggest that FMDV Lbpro-mediated targeting of STAT1/2 may reveal a novel mechanism for viral immune evasion.

Structural basis for IFN antagonism by human respiratory syncytial virus nonstructural protein 2.

Human respiratory syncytial virus (RSV) nonstructural protein 2 (NS2) inhibits host interferon (IFN) responses stimulated by RSV infection by targeting early steps in the IFN-signaling pathway. But the molecular mechanisms related to how NS2 regulates these processes remain incompletely understood. To address this gap, here we solved the X-ray crystal structure of NS2. This structure revealed a unique fold that is distinct from other known viral IFN antagonists, including RSV NS1. We also show that NS2 directly interacts with an inactive conformation of the RIG-I-like receptors (RLRs) RIG-I and MDA5. NS2 binding prevents RLR ubiquitination, a process critical for prolonged activation of downstream signaling. Structural analysis, including by hydrogen-deuterium exchange coupled to mass spectrometry, revealed that the N terminus of NS2 is essential for binding to the RIG-I caspase activation and recruitment domains. N-terminal mutations significantly diminish RIG-I interactions and result in increased IFNß messenger RNA levels. Collectively, our studies uncover a previously unappreciated regulatory mechanism by which NS2 further modulates host responses and define an approach for targeting host responses.

Targeting BCAA metabolism to potentiate metformin's therapeutic efficacy in the treatment of diabetes in mice.

AIMS/HYPOTHESIS: An increasing body of evidence has shown that the catabolism of branched-chain amino acids (BCAAs; leucine, isoleucine and valine) is impaired in obese animals and humans, contributing to the development of insulin resistance and type 2 diabetes. Promoting BCAA catabolism benefits glycaemic control. It remains unclear whether BCAA catabolism plays a role in the therapeutic efficacy of currently used glucose-lowering drugs such as metformin. METHODS: Mice were treated with vehicle or metformin (250 mg/kg per day) for more than 4 weeks to investigate the effects of metformin in vivo. In vitro, primary mouse hepatocytes and HepG2 cells were treated with 2 mmol/l metformin. The therapeutic efficacy of metformin in the treatment of type 2 diabetes was assessed in genetically obese (ob/ob) mice and high-fat-diet-induced obese (DIO) mice. Enhancing BCAA catabolism was achieved with a pharmacological agent, 3,6-dichlorobenzo[b]thiophene-2-carboxylic acid (BT2). The ob/ob mice were treated with a low-BCAA diet or intermittent protein restriction (IPR) to reduce BCAA nutritional intake. RESULTS: Metformin unexpectedly inhibited the catabolism of BCAAs in obese mice, resulting in an elevation of BCAA abundance. AMP-activated protein kinase (AMPK) mediated the impact of metformin on BCAA catabolism in hepatocytes. Importantly, enhancing BCAA catabolism via a pharmacological agent BT2 significantly potentiated the glucose-lowering effect of metformin while decreasing circulating BCAA levels in ob/ob and DIO mice. Similar outcomes were achieved by a nutritional approach of reducing BCAA intake. IPR also effectively reduced the circulating BCAA abundance and enhanced metformin's glucose-lowering effect in ob/ob mice. BT2 and IPR treatments reduced the expression of fructose-1,6-bisphosphatase 1, a rate-limiting enzyme in gluconeogenesis, in the kidney but not liver, indicating the involvement of renal gluconeogenesis. CONCLUSIONS/INTERPRETATION: Metformin self-limits its therapeutic efficacy in the treatment of type 2 diabetes by triggering the suppression of BCAA catabolism. Enhancing BCAA catabolism pharmacologically or reducing BCAA intake nutritionally potentiates the glucose-lowering effect of metformin. These data highlight the nutritional impact of protein on metformin's therapeutic efficacy and provide new strategies targeting BCAA metabolism to improve metformin's effects on the clinical outcome in diabetes.

Combinational Deletions of MGF360-9L and MGF505-7R Attenuated Highly Virulent African Swine Fever Virus and Conferred Protection against Homologous Challenge.

Multigene family (MGF) gene products are increasingly reported to be implicated in African swine fever virus (ASFV) virulence and attenuation of host defenses, among which the MGF360-9L and MGF505-7R gene products are characterized by convergent but distinct mechanisms of immune evasion. Herein, a recombinant ASFV mutant, ASFV-Δ9L/Δ7R, bearing combinational deletions of MGF360-9L and MGF505-7R, was constructed from the highly virulent ASFV strain CN/GS/2018 of genotype II that is currently circulating in China. Pigs inoculated intramuscularly with 104 50% hemadsorption doses (HAD50) of the mutant remained clinically healthy without any serious side effects. Importantly, in a virulence challenge, all four within-pen contact pigs demonstrated clinical signs and pathological findings consistent with ASF. In contrast, vaccinated pigs (5/6) were protected and clinical indicators tended to be normal, accompanied by extensive tissue repairs. Similar to most viral infections, innate immunity and both humoral and cellular immune responses appeared to be vital for protection. Notably, transcriptome sequencing (RNA-seq) and quantitative PCR (qPCR) analysis revealed a regulatory function of the mutant in dramatic and sustained expression of type I/III interferons and inflammatory and innate immune genes in vitro. Furthermore, infection with the mutant elicited an early and robust p30-specific IgG response, which coincided and was strongly correlated with the protective efficacy. Analysis of the cellular response revealed a strong ASFV-specific interferon gamma (IFN-γ) response and immunostaining of CD4+ T cells coupled with a high level of CD163+ macrophage infiltration in spleens of vaccinated pigs. Our study identifies a new mechanism of immunological regulation by ASFV MGFs that rationalizes the design of live attenuated vaccine for implementation of improved control strategies to eradicate ASFV. IMPORTANCE Currently, the deficiency in commercially available vaccines or therapeutic options against African swine fever constitutes a matter of major concern in the swine industry globally. Here, we report the design and construction of a recombinant ASFV mutant harboring combinational deletions of interferon inhibitors MGF360-9L and MGF505-7R based on a genotype II ASFV CN/GS/2018 strain currently circulating in China. The mutant was completely attenuated when inoculated at a high dose of 104 HAD50. In the virulence challenge with homologous virus, sterile immunity was achieved, demonstrating the mutant's potential as a promising vaccine candidate. This sufficiency of effectiveness supports the claim that this live attenuated virus may be a viable vaccine option with which to fight ASF.

FMDV 3A Antagonizes the Effect of ANXA1 to Positively Modulate Viral Replication.

The RIG-I-like receptor signaling pathway is crucial for producing type I interferon (IFN-I) against RNA viruses. The present study observed that viral infection increased annexin-A1 (ANXA1) expression, and ANXA1 then promoted RNA virus-induced IFN-I production. Compared to ANXA1 wild-type cells, ANXA1-/- knockout cells showed IFN-ß production decreasing after viral stimulation. RNA virus stimulation induced ANXA1 to regulate IFN-ß production through the TBK1-IRF3 axis but not through the NF-κB axis. ANXA1 also interacted with JAK1 and STAT1 to increase signal transduction induced by IFN-ß or IFN-γ. We assessed the effect of ANXA1 on the replication of foot-and-mouth disease virus (FMDV) and found that ANXA1 inhibits FMDV replication dependent on IFN-I production. FMDV 3A plays critical roles in viral replication and host range. The results showed that FMDV 3A interacts with ANXA1 to inhibit its ability to promote IFN-ß production. We also demonstrated that FMDV 3A inhibits the formation of ANXA1-TBK1 complex. These results indicate that ANXA1 positively regulates RNA virus-stimulated IFN-ß production and FMDV 3A antagonizes ANXA1-promoted IFN-ß production to modulate viral replication. IMPORTANCE FMDV is a pathogen that causes one of the world's most destructive and highly contagious animal diseases. The FMDV 3A protein plays a critical role in viral replication and host range. Although 3A is one of the viral proteins that influences FMDV virulence, its underlying mechanisms remain unclear. ANXA1 is involved in immune activation against pathogens. The present study demonstrated that FMDV increases ANXA1 expression, while ANXA1 inhibits FMDV replication. The results also showed that ANXA1 promotes RNA virus-induced IFN-I production through the IRF3 axis at VISA and TBK1 levels. ANXA1 was also found to interact with JAK1 and STAT1 to strengthen signal transduction induced by IFN-ß and IFN-γ. 3A interacted with ANXA1 to inhibit ANXA1-TBK1 complex formation, thereby antagonizing the inhibitory effect of ANXA1 on FMDV replication. This study helps to elucidate the mechanism underlying the effect of the 3A protein on FMDV replication.

The relationship between screen time and attention deficit/hyperactivity disorder in Chinese preschool children under the multichild policy: a cross-sectional survey.

BACKGROUND: Long screen time has become a public health problem that cannot be ignored. The association between screen time and attention-deficit/hyperactivity disorder (ADHD) in preschool children has received widespread attention. METHODS: A questionnaire was used to survey 2452 people. ADHD symptoms were assessed by the Conners Child Behavior Scale. Considering that the ADHD symptoms of boys and girls might be different, we stratified the data by gender. Logistic regression model was used for regression analysis. To exclude the influence of multichild family and obesity level, we also conducted a sensitivity analysis. P values were two-tailed with a significance level at 0.05. RESULTS: The results showed that the association between screen time and ADHD symptoms in preschool children was significant (OR = 1.826, 95%CI: 1.032, 3.232). After grouping the genders, the correlation was not significant. There was an association between screen time and ADHD symptoms in children from families with multiple children. However, after excluding overweight and obese children from the overall population, the association between screen time and ADHD symptoms did not have statistical significance. CONCLUSIONS: The issue of screen time for preschoolers needs to be taken seriously. Although the results indicate a significant correlation between screen time and ADHD symptoms, clearer evidence is needed to provide recommendations to policy makers.

A model to evaluate ozone distribution and reaction byproducts in aircraft cabin environments.

Ozone and byproducts of ozone-initiated reactions are among the primary pollutants in aircraft cabins. However, investigations of the spatial distribution and reaction mechanisms of these pollutants are insufficient. This study established a computational fluid dynamics-based model to evaluate ozone and byproduct distribution, considering ozone reactions in air, adsorption onto surfaces, and byproduct desorption from surfaces. The model was implemented in an authentic single-aisle aircraft cabin and validated by measurements recorded during the aircraft cruise phase. Ozone concentrations in the supply air-dominated area were approximately 50% higher than that in the passenger breathing zone, suggesting that human surfaces represent a significant ozone sink. The deposition velocity onto human bodies was 21.83 m/h, surpassing 3.97 m/h on other cabin interior surface areas. Our model provides a mechanistic tool to analyze ozone and byproduct concentration distributions, which would be useful for assessing passenger health risks and for developing strategies for healthier aircraft cabin environments.

Rapid identification of candidate genes controlling male-sterility in Foxtail millet (Setaria italica).

Photo-(thermo-) sensitive genic male-sterile line is the key component of two-line hybridization system in foxtail millet (Setaria italica), but the genetic basis of male sterility in most male-sterile lines is still unclear. In the present study, a large F2 population was developed derived from a cross between the photo-(thermo-) sensitive male-sterile line A2 and the fertile-line 1484-5. Thirty plants with extreme high and extreme low fertility were selected from the population to construct a sterile DNA pool and a fertile DNA pool, respectively. Sequencing both DNA pools and data analysis revealed that two QTLs conferred male-sterility, qSiMS6.1 with a major effect and qSiMS6.2 with a minor effect, on chromosome 6. Both QTLs exhibited complete dominance. The major QTL, qSiMS6.1, was delimited to a 186-kb interval between the markers SiM20 and SiM9 by the joint analysis of QTL-seq and QTL mapping with SSR and structure variation markers. Millet_GLEAN_10020454 in this region is the most likely candidate gene for qSiMS6.1 since it is predicted to encode a male-sterile 5 like protein. These results lay a solid foundation for qSiMS6.1 cloning and provided gene resources for breeding new male-sterile lines. Supplementary Information: The online version contains supplementary material available at 10.1007/s11032-021-01269-2.

Carbon dioxide in passenger cabins: Spatial temporal characteristics and 30-year trends.

Carbon dioxide (CO2 ) is an important environmental parameter in aircraft cabins. To understand the most recent, real-time CO2 concentration levels and their key influencing factors in aircraft cabins, we conducted in-flight measurements of 52 randomly selected commercial flights with different aircraft types and durations from August 2017 to August 2019. The spatial temporal characteristics of CO2 concentrations on board were analyzed and summarized. For the flight time scale, the CO2 concentrations during the boarding phase (1680 ± 558 ppmv) were notably higher than that in other phases, whereas the condition of the cruising phase was the lowest in most flights. The flight average CO2 concentrations of the cruising phase were 1253 ± 164 ppmv, and the corresponding estimated outside airflow rates were 6.2 ± 1.3 L/s/p in the economy class across all flights. Single-aisle and twin-aisle flights did not show noticeable differences for the same phases. Relatively uniform CO2 concentrations were observed at different positions of the same class. By comparing the results of this study with those previously reported, CO2 concentrations showed a slightly decreasing trend over the last 30 years. This suggested a slightly increased ventilation rate and potentially superior air quality on board.

Influencing factors of carbonyl compounds and other VOCs in commercial airliner cabins: On-board investigation of 56 flights.

Volatile organic compounds (VOCs) as a non-negligible aircraft cabin air quality (CAQ) factor influence the health and comfort of passengers and crew members. On-board measurements of carbonyls (short-chain (C1 -C6 )) and other volatile organic compounds (VOCs, long-chain (C6 -C16 )) with a total of 350 samples were conducted in 56 commercial airliner cabins covering 8 aircraft models in this study. The mean concentration for each individual carbonyl compound was between 0.3 and 8.3 µg/m3 (except for acrolein & acetone, average = 20.7 µg/m3 ) similar to the mean concentrations of other highly detected VOCs (long-chain (C6 -C16 ), 97% of which ranged in 0-10 µg/m3 ) in aircraft cabins. Formaldehyde concentrations in flights were significantly lower than in residential buildings, where construction materials are known formaldehyde sources. Acetone is a VOC emitted by humans, and its concentration in flights was similar to that in other high-occupant density transportation vehicles. The variation of VOC concentrations in different flight phases of long-haul flights was the same as that of CO2 concentration except for the meal phase, which indicates the importance of cabin ventilation in diluting the gaseous contaminants, while the sustained and slow growth of the VOC concentrations during the cruising phase in short-haul flights indicated that the ventilation could not adequately dilute the emission of VOCs. For the different categories of VOCs, the mean concentration during the cruising phase of benzene series, aldehydes, alkanes, other VOCs (detection rate > 50%), and carbonyls in long-haul flights was 44.2 µg/m3 , 17.9 µg/m3 , 18.6 µg/m3 , 31.5 µg/m3 , and 20.4 µg/m3  lower than those in short-haul flights, respectively. Carbonyls and d-limonene showed a significant correlation with meal service (p < 0.05). Unlike the newly decorated rooms or new vehicles, the inner materials were not the major emission sources in aircraft cabins. Practical Implications. The on-board measurements of 56 flights enrich the VOC database of cabin environment, especially for carbonyls. The literature review of carbonyls in the past 20 years contributes to the understanding the current status of cabin air quality (CAQ). The analysis of VOC concentration variation for different flight phases, flight duration, and aircraft age lays a foundation for exploring effective control methods, including ventilation and purification for cabin VOC pollution. The enriched VOC data is helpful to explore the key VOCs of aircraft cabin environment and to evaluate the acute/chronic health exposure risk of pollutants for passengers and crew members.

Association between maternal vitamin D status with pregnancy outcomes and offspring growth in a population of Wuxi, China.

BACKGROUND AND OBJECTIVES: The role of maternal vitamin D in infantile growth remains unclear. METHODS AND STUDY DESIGN: Serum 25-hydroxyvitamin D [25(OH)D] concentrations were examined for pregnancies who visited the Affiliated Wuxi Maternity and Child Health Care Hospital of Nanjing Medical University from January 2016 to December 2017. Anthropometric measurements of corresponding offspring were performed from birth to 2 to 3 years old. Infantile body mass index (BMI) was transformed into age-, sex- and height- normalized z scores, and Latent Class Growth Mixture (LCGM) model was used to identify trajectories of BMI-Z. RESULTS: Among the 329 included pregnancy women, 109 (33.13 %), 190 (57.75%) and 30 (9.12%) were defined as vitamin D deficiency [25(OH)D <30 nmol/L], insufficiency [30 nmol/L≤25(OH)D<50 nmol/L] and sufficiency [25(OH)D ≥50 nmol/L], respectively. When compared with vitamin D sufficiency, maternal vitamin D deficiency was not associated with preterm birth [odds ratio (OR)=2.69, 95% confidence interval (95% CI)=0.57-12.80], small for gestation age (OR=0.99, 95% CI=0.29-3.46), and low birth weight (OR=1.69, 95% CI=0.34-8.51). Similarly, no significant relationships were found between maternal vitamin D concentrations and anthropometric indices (such as weight, length, BMI) during 0 to 3 years old. Furthermore, LCGM model identified two patterns of offspring growth: stable moderate BMI-Z and early transient BMI-Z groups. Maternal vitamin D levels were higher in the former group than the latter (p=0.037); however, maternal vitamin D status appeared to be unrelated with offspring BMI-Z trajectories in multivariable logistic regression models. CONCLUSIONS: Maternal vitamin D deficiency may not be related to adverse pregnancy outcomes as well as offspring growth.

[Analysis of FGD1 gene variant in a child with Aarskog-Scott syndrome].

OBJECTIVE: To detect pathogenic variant of the FGD1 gene in a boy with Aarskog-Scott syndrome. METHODS: Genetic variant was detected by high-throughput sequencing. Suspected variant was verified by Sanger sequencing. The nature and impact of the candidate variant were predicted by bioinformatic analysis. RESULTS: The child was found to harbor a novel c.1906C>T hemizygous variant of the FGD1 gene, which has led to conversion of Arginine to Tryptophane at codon 636(p.Arg636Trp). The same variant was found in his mother but not father. Based on the American College of Medical Genetics and Genomics guidelines, the c.1906C>T variant of FGD1 gene was predicted to be likely pathogenic(PM1+PM2+PM5+PP2+PP3+PP4). CONCLUSION: The novel c.1906C>T variant of the FGD1 gene may underlay the Aarskog-Scott syndrome in this child. Above finding has enabled diagnosis for the boy.

A field investigation of the thermal environment and adaptive thermal behavior in bedrooms in different climate regions in China.

Sleep thermal environments substantially impact sleep quality. To study the sleep thermal environment and thermal comfort in China, this study carried out on-site monitoring of thermal environmental parameters in peoples' homes, including 166 households in five climate zones, for one year. A questionnaire survey on sleep thermal comfort and adaptive behavior was also conducted. The results showed that the indoor temperature for sleep in northern China was more than 4°C higher than that in southern China in winter, while the indoor temperatures for sleep were similar in summer. Furthermore, 70% of people were satisfied with their sleep thermal environment. Due to the use of air conditioning and window opening in various areas in summer, people were satisfied with their sleep thermal environments. Due to the lack of central heating in the southern region in winter, people feel cold and their sleep thermal environment needs further improvement. The bedding insulation in summer and winter in northern China was 1.83clo and 2.67clo, respectively, and in southern China was 2.21clo and 3.17clo, respectively. Both northern China and southern China used air conditioning only in summer. People in southern China opened their windows all year, while those in northern China opened their windows during the summer and transitional periods.

Study on the Adsorption Behavior between an Imidazolium Ionic Liquid and Na-Montmorillonite.

Interactions between 1-butyl-3-methylimidazolium tetrafluoroborate (IL), an ionic liquid, and Na-montmorillonite (Na-MMT) were studied under different kinetic conditions to investigate the adsorption behavior of IL by Na-MMT. The adsorption of IL by Na-MMT was rapid, with a fast rate, reaching a capacity of 0.43 mmol/g, lower than Na-MMT's cation exchange capacity (CEC) of 0.90 mmol/g. Meanwhile, the highest adsorption rate occurred at the IL concentration of 1000 mg/L. The exchangeable cation of Na-MMT could not be completely substituted by the cation group of IL regardless of the IL concentration. Stoichiometric desorption experiments confirmed that the cation exchange was the dominating adsorption mechanism for the IL adsorption by Na-MMT. The pH value of the solution between 2 and 11 had a negligible effect on the adsorption amount of IL by Na-MMT. The cation group of IL interacted into the interlayer of Na-MMT successfully, resulting in the change in the wettability of Na-MMT. A bilayer formation of the cationic group should occur in the interlayer of the modified Na-MMT and the configuration of IL was dependent on the adsorption amount of IL. Furthermore, the thermal stability of the modified Na-MMT was also dependent on the adsorption amount of IL.

Rapid and sensitive detection of porcine epidemic diarrhea virus by reverse transcription loop-mediated isothermal amplification combined with a vertical flow visualization strip.

Porcine epidemic diarrhea virus (PEDV) is an important pathogen that causes vomiting, diarrhea, and dehydration, leading to serious damage to the swine industry worldwide. The establishment of effective diagnostic methods is imperative. However, traditional methods are often unsuitable. In this study, reverse transcription loop-mediated isothermal amplification (RT-LAMP) was combined with a vertical flow (VF) nucleic acid detection strip to detect PEDV. Parameters that affect the RT-LAMP reaction were optimized. The RT-LAMP-VF assay that we established was performed at 62 °C for 40 min, and then directly evaluated on the VF visualization strip cassette. The method demonstrated high specificity for PEDV. The detection limit was 10 pg of ribonucleic acid, consistent with RT-PCR, RT-LAMP detected products on agarose gels and by direct calcein fluorescence. Application of this method to clinical samples yielded a positivity rate that was comparable to that obtained for RT-PCR. This technique saves time and is efficient, and is thus expected to be useful for the diagnosis of PEDV infection in the field.

Mutational spectrum of phenylketonuria in Jiangsu province.

UNLABELLED: Phenylketonuria (PKU) is caused by variants in the phenylalanine hydroxylase (PAH) gene. We systematically investigated all 13 exons of the PAH gene and their flanking introns in 31 unrelated patients and their parents using next-generation sequencing (NGS). A total of 33 different variants were identified in 58 of 62 mutant PAH alleles. The prevalent variants with a relative frequency of 5 % or more were c.721C > T, c.1068C > A, c.611A > G, c.1197A > T, c.728G > A, c.331C > T, and c.442-1G > A. One novel variant was identified in this study-c.699C > G. We studied genotype-phenotype correlations using the Guldberg arbitrary value (AV) system, which revealed a consistency rate of 38 % (8/21) among the 21 predicted phenotypes. The genotype-based prediction of BH4 responsiveness was also evaluated, and 14 patients (45.2 %) were predicted to be BH4 responsive. CONCLUSION: This study presents the spectrum of PAH variants in Jiangsu province. The information obtained from the genotype-based prediction of BH4 responsiveness might be used for the rational selection of candidates for BH4 testing. WHAT IS KNOWN: • Phenylketonuria (PKU) is caused by variants in the phenylalanine hydroxylase (PAH) gene. • The spectrum of PAH variants in different Chinese populations has been reported. What is new: • This is the first report on the spectrum of PAH variants in Jiangsu province. • This study identified one novel PAH variant-c.699C>G-and and tries to show a genotype-phenotype relationship also regarding BH4-responsiveness.

Experimental study of gaseous and particulate contaminants distribution in an aircraft cabin.

The environment of the aircraft cabin greatly influences the comfort and health of passengers and crew members. Contaminant transport has a strong effect on disease spreading in the cabin environment. To obtain the complex cabin contaminant distribution fields accurately and completely, which is also essential to provide solid and precise data for computational fluid dynamics (CFD) model validation, this paper aimed to investigate and improve the method for simultaneous particle and gaseous contaminant fields measurement. The experiment was conducted in a functional MD-82 aircraft. Sulfur hexafluoride (SF6) was used as tracer gas, and Di-Ethyl-Hexyl-Sebacat (DEHS) was used as particulate contaminant. The whole measurement was completed in a part of the economy-class cabin without heating manikins or occupied with heating manikins. The experimental method, in terms of pollutant source setting, sampling points and schedule, was investigated. Statistical analysis showed that appropriately modified sampling grid was able to provide reasonable data. A small difference in the source locations can lead to a significant difference in cabin contaminant fields. And the relationship between gaseous and particulate pollutant transport was also discussed through tracking behavior analysis.

[Preparation of a mouse monoclonal antibody against the NS1 protein of respiratory syncytial virus].

The aim of this study was to prepare a mouse monoclonal antibody against the nonstructural protein 1 (NS1) of respiratory syncytial virus (RSV) to analyze its expression and distribution during transfection and infection. Additionally, we aimed to evaluate the antibody's application in immunoprecipitation assay. Firstly, the NS1 gene fragment was cloned into a prokaryotic plasmid and expressed in Escherichia coli. The resulting NS1 protein was then purified by affinity chromatography, and used to immunize the BALB/c mice. Subsequently, hybridoma cells capable of stably secreting the NS1 monoclonal antibody were selected using indirect enzyme linked immunosorbent assay (ELISA). This monoclonal antibody was employed in both indirect immunofluorescence assay (IFA) and Western blotting to analyze the expression and distribution of RSV NS1 in overexpressed and infected cells. Finally, the reliability of this monoclonal antibody was evaluated through the immunoprecipitation assay. The results showed that the RSV NS1 protein was successfully expressed and purified. Following immunization of mice with this protein, we obtained a highly specific RSV NS1 monoclonal antibody, which belonged to the IgG1 subtype with an antibody titer of 1:15 360 000. Using this monoclonal antibody, the RSV NS1 protein was identified in both transfected and infected cells. The IFA results revealed predominant distribution of NS1 in the cytoplasm and nucleus. Moreover, we confirmed that this monoclonal antibody could effectively bind specifically to NS1 protein in cell lysates, making it suitable as a capture antibody in immunoprecipitation assay. In conclusion, our study successfully achieved production of the RSV NS1 protein through a prokaryotic expression system and prepared a specific monoclonal antibody against NS1. This antibody demonstrates the ability to specifically identify the NS1 protein and can be used in the immunoprecipitation assay, thereby laying a foundation for the functional studies of the NS1 protein.

The multiple roles of viral 3Dpol protein in picornavirus infections.

The Picornaviridae are a large group of positive-sense, single-stranded RNA viruses, and most research has focused on the Enterovirus genus, given they present a severe health risk to humans. Other picornaviruses, such as foot-and-mouth disease virus (FMDV) and senecavirus A (SVA), affect agricultural production with high animal mortality to cause huge economic losses. The 3Dpol protein of picornaviruses is widely known to be used for genome replication; however, a growing number of studies have demonstrated its non-polymerase roles, including modulation of host cell biological processes, viral replication complex assembly and localization, autophagy, and innate immune responses. Currently, there is no effective vaccine to control picornavirus diseases widely, and clinical therapeutic strategies have limited efficiency in combating infections. Many efforts have been made to develop different types of drugs to prohibit virus survival; the most important target for drug development is the virus polymerase, a necessary element for virus replication. For picornaviruses, there are also active efforts in targeted 3Dpol drug development. This paper reviews the interaction of 3Dpol proteins with the host and the progress of drug development targeting 3Dpol.