Parainfluenza Virus Type Research Articles

P-1170. Impact of Healthcare-associated Respiratory Virus Infections in Children

Abstract Background Respiratory viruses can spread within healthcare facilities despite stringent infection prevention measures. Although healthcare-associated respiratory virus infections (HARVI) pose a significant risk to children, pediatric HARVI are understudied. The present study aimed to evaluate the impact of HARVI on children following the SARS-CoV-2 pandemic at a tertiary children’s hospital. Methods This retrospective study enrolled pediatric inpatients aged ≤18 years who were hospitalized at Tokyo Metropolitan Children's Medical Center between April 2020 and March 2024. HARVI was defined as a viral infection with a new onset of respiratory symptoms past the incubation period following hospitalization. The causative virus was detected using multiplex PCR (FilmArray Respiratory Panel 2.1, BioFire). Patients with other infections were excluded. The primary outcome was the 30-day mortality rate. The secondary outcomes included PICU admission, escalation of respiratory support, and the postponement of discharge, planned surgery or chemotherapy. Results In total, 265 patients with HARVI were enrolled. Male patients comprised 31% of the cohort, and the median age was 31 (IQR [interquartile range]: 12-77) months. Underlying neurologic, cardiac, and congenital or genetic disorders had in 42%, 28%, and 24% of the cohort, respectively. The respiratory viruses detected were the rhinovirus/enterovirus (44%), parainfluenza virus type 3 (15%), and HKU1, NL63, 229E and OC43 coronaviruses (8%). The 30-day mortality rate was 1% in three patients with parainfluenza virus type 3, RSV, and SARS-CoV-2. The PICU admission rate was 2%. The rate of respiratory support escalation was 17%. Postponement of discharge was required in 7% of the cohort for a median period of seven (IQR: 5-8) days. Postponement of surgery was required in 3% of the cohort for a median period of 46 (IQR: 21-82) days. There was no postponement of chemotherapy. Conclusion Pediatric HARVI was associated with mortality and additional medical care. Therefore, strict infection prevention measures are essential. Disclosures Hanako Funakoshi, MD, Pfizer Japan: Husband is an employee.

P-703. Manifestations of parainfluenza type 4 infections and viral loads in children

Abstract Background Although parainfluenza virus type 4 (PIV-4) typically causes mild respiratory illnesses in children, it can cause severe diseases occasionally. It is not known Whether viral load of PIV-4 could correlate with severity. This study aimed to assess the relationship between PIV-4 viral load and disease’s severity in children. Viral load and severity This figure shows the relationship between severity of illness and viral load.There was no correlation between viral load and severity. Methods A retrospective study at Tokyo Metropolitan Children's Medical Center from April to October 2023 screened hospitalized children using multiplex PCR. PIV-4 positive samples were assayed for viral load of all detected viruses using real-time PCR. Cases with PIV-4 were included if PIV-4 was only detected or the viral load of PIV-4 was higher than other co-detected viruses. Demographic data, underlying diseases, laboratory, co-infections, diagnoses, and outcomes were collected from electrical charts. Cases with PIV-4 were categorized into mild cases with no oxygen demand, moderate cases with low-flow oxygen demand and severe cases with high flow nasal cannula oxygen or non-invasive/invasive mechanical ventilation or PICU admission.The viral loads of PIV-4 and number of co-infected virus were compared among severity groups. Characteristics, Symptoms and Laboratory Data of PIV-4 positive patients Results Nasal swab samples were collected from 2884 cases. PIV-4 was detected in 185 samples. As 108 cases were excluded for higher viral loads of other viruses, 77 cases were included for analysis. The severity of PIV-4 comprised 37 mild, 24 moderate, and 16 severe cases. The rate of lower respiratory tract infections in mild, moderate and severe cases were 8%, 96%, and 75%, respectively. There was only one PIV-4-related mortality at 28 days. Co-detection with rhinovirus/enterovirus was the most frequent. Median viral loads (log10 copies/mL) in mild, moderate and severe cases were 5.98 (IQR 3.58-6.89), 6.09 (IQR 4.93-6.66) and 5.98 (IQR 3.57-7.03), respectively. No significant correlation was found between viral load and severity, but severity correlated with the number of viral co-infections. Treatment and Outcome of PIV-4 positive patients Conclusion Moderate or severe Pediatric PIV-4 infections primarily show as lower respiratory tract infections, with no relation between viral load and illness severity, though the number of co-infected virus may play a role. Severity and co-infection viruses Disclosures All Authors: No reported disclosures

P-1207. Resurgence of Respiratory Viral Infections Among Children Hospitalized in a Tertiary Care Hospital in Thailand After the COVID-19 Pandemic

Abstract Background To demonstrate the epidemiological and clinical characteristics of respiratory viral infections among children hospitalized in a tertiary care hospital after COVID-19 pandemic. Methods A cross-sectional study was conducted among children aged 1 month to 15 years who were admitted to the Chiang Mai University Hospital, Thailand due to non-COVID-19 respiratory tract infections (RTIs) after COVID-19 pandemic (Oct 2022 to Sep 2023). Nasopharyngeal specimens were tested for 16 respiratory viruses (RV: influenza A [IAV] and B [IBV] viruses; respiratory syncytial virus [RSV]; parainfluenza virus types 1-4; rhinovirus; enterovirus; human metapneumovirus; human coronaviruses 229, NL63, OC43, HKU1; bocavirus; human adenovirus), using multiplex RT-PCR. The prevalence each RV was demonstrated. Clinical parameters, treatments, and outcomes of children were compared by type of RV. In addition, the prevalence of IAV, IBV and RSV were compared across 3 periods: pre (Jan 2016 to Dec 2019), during (Jan 2020 to Sep 2022), and post COVID-19 pandemic. Results During the study period, 400 children were enrolled; median age 2.9 (IQR 1.5-4.8) years, 59% male, 46% had underlying diseases. On admission, 80% had pneumonia and 5% were admitted to ICUs. Overall, 341 children (85%) were tested positive for ≥ 1 RV (68% single and 17% multiple infections). The prevalence of each RV after the COVID-19 pandemic was shown in Table 1. Compared with other viruses, children with IAV and IVB (n=21) had the highest proportion of fever (100%) and sore throat (19%), whereas those with RSV (n=65) had the highest proportion of oxygen therapy (97%) and bronchodilator (86%) requirement, and those with enterovirus (n=23) had the highest proportion of tachypnea (44%), ICU admission/transfer (22%), ventilator use (22%), and death (4%) (Table 2). Notably, the prevalence of IAV, IBV and RSV significantly declined during the COVID-19 pandemic, and sharply rebounded after the pandemic, particularly for RSV which the prevalence was significantly higher than pre COVID-19 pandemic phase (Figure 1). Conclusion COVID-19 pandemic significantly changed the epidemiology of respiratory viral infections in our setting. Active surveillance is needed to better understand the dynamics and seasonal patterns of RV after the pandemic. Disclosures All Authors: No reported disclosures

P-672. Coinfections with Respiratory Syncytial Virus (RSV) in a Cohort of Adults with Pre-Existing Comorbidities in Wisconsin from 2015-16 through 2019-20

Abstract Background Current evidence suggests that coinfections with respiratory syncytial virus (RSV) may affect symptom and disease severity during acute respiratory infection (ARI). However, this data is largely observed in children, with limited data regarding RSV coinfections in adults with pre-existing comorbidities.Table 1.Demographic and clinical characteristics among high-risk adults with RSV coinfection and RSV single infection from 2015-16 through 2019-20 respiratory virus seasons. Methods We conducted a retrospective cohort study, using existing data and respiratory specimens from community-dwelling adult participants in a test-negative study of influenza vaccine effectiveness. The study was conducted in an outpatient setting in Wisconsin from 2015-2016 through 2019-2020. Participants included in this analysis had ≥1 pre-existing comorbidity (Table 1). Residual respiratory specimens were retested using a PCR-based multiplex panel for adenovirus (HAdV), seasonal coronaviruses (HCoV), human metapneumovirus, rhinovirus/enterovirus (HRV), influenza virus (IFV) (influenza A H1pdm09, H3, and B), human parainfluenza virus types 1-4, RSV A and B. RSV coinfection was defined as the detection of two or more pathogens, one of which was RSV A or B and the other non-RSV. We used chi-square tests to compare demographic and clinical characteristics between those with RSV coinfection vs. RSV single infection. Results Of 3601 respiratory samples from high-risk adults tested, RSV coinfection was detected in 18 (0.5%) participants and RSV single infection in 285 (7.9%). RSV coinfections occurred with HRV (8; 44.4%); IFV (6; 33.3%); HCoV (5; 27.8%); and HAdV (1; 5.6%), and was highest among those aged 18-49 years (9; 50%). We did not observe statistically significant differences between coinfections vs. single infections by presence of upper or lower respiratory symptoms, or by high-risk conditions (Table 1). Hospitalization occurred only in individuals with RSV single infections (10, 3.5%). Conclusion RSV viral co-infections were detected in 0.5% of 3601 respiratory samples of high-risk adults, including coinfections with HRV, IFV, HCoV and HAdV. We did not observe significant differences in symptoms between those with RSV coinfection vs single infection, however, hospitalization occurred only in individuals with RSV single infection. Limitations of this analysis include its sample size and the restrictions of multiplex assay sensitivity and specificity. Disclosures Oluwakemi Alonge, MPH, CPH, CSL Seqirus: Grant/Research Support|GSK Inc.: Grant/Research Support Maria Sundaram, PhD, MSPH, GSK Inc.: Grant/Research Support|Pfizer Inc.: Grant/Research Support Huong Nguyen, PhD, MPH, CSL Seqirus: Advisor/Consultant|CSL Seqirus: Grant/Research Support|GSK: Grant/Research Support|ModernaTX: Advisor/Consultant|ModernaTX: Grant/Research Support Jennifer P. King, MPH, GSK Inc.: Grant/Research Support|ModernaTX: Grant/Research Support Elisha Stefanski, BS, CSL Seqirus: Grant/Research Support|GSK Inc.: Grant/Research Support|ModernaTX: Grant/Research Support Pouya Saeedi, PhD., GSK: Employment Yves Brabant, MEng, GSK: Employed Jean-Yves Pirçon, PhD, GSK Inc.: Employment

The Combination of GS-441524 (Remdesivir) and Ribavirin Results in a Potent Antiviral Effect Against Human Parainfluenza Virus 3 Infection in Human Airway Epithelial Cell Cultures and in a Mouse Infection Model

Human parainfluenza virus type 3 (HPIV-3) can cause severe respiratory diseases, particularly in young children, the elderly and immunocompromised. There are no approved antiviral drugs against this virus. We report that the combination of ribavirin with either remdesivir or its parent nucleoside GS-441524 results in a pronounced antiviral effect against HPIV-3 in LLC-MK2 cells and in human airway epithelial cells grown at the air–liquid interface. In AG129 mice intranasally inoculated with HPIV-3, the combined treatment with ribavirin and GS-441524 decreased infectious viral lung titers by >2.5 log10 to undetectable levels in 4 out of 11 mice and by 1.6 log10 in the remaining 7 mice as compared with the vehicle. The lungs of all mice that received the combined treatment appeared histologically normal or virtually normal, whereas 8 of 11 vehicle-treated mice presented with bronchopneumonia. By contrast, ribavirin alone did not result in a reduction in infectious viral lung titers; GS-441524 alone reduced infectious viral lung titers by 1.2 log10. Moreover, several mice in the single-treatment groups exhibited severe lung pathology. These findings may warrant exploring this combination in patients with severe HPIV-3 infections and possibly also against infections with other viruses that are susceptible in vitro to these two drugs.

Single-cycle parainfluenza virus type 5 vectors for producing recombinant proteins, including a humanized anti-V5 tag antibody.

Parainfluenza virus type 5 (PIV5) can cause either persistent or acute/lytic infections in a wide range of mammalian tissue culture cells. Here, we have generated PIV5 fusion (F)-expressing helper cell lines that support the replication of F-deleted viruses. As proof of the principle that F-deleted single-cycle infectious viruses can be used as safe and efficient expression vectors, we have cloned and expressed a humanized (Hu) version of the mouse anti-V5 tag antibody (clone SV5-Pk1). We show that multiple different cell lines can be infected and express high levels of the Hu anti-V5 antibody, with Chinese hamster ovary cells expressing 20-50 mg l-1 after 5 days when cells were grown to a density of ~1×106 cells per millilitre at the time of infection. We suggest that PIV5-based vectors may be further developed to produce recombinant proteins both in vitro and in vivo.

Temporal Trends in Respiratory Infection Epidemics Among Pediatric Inpatients Throughout the Course of the COVID-19 Pandemic From 2018 to 2023 in Fukushima Prefecture, Japan.

Nonpharmaceutical interventions for coronavirus disease (COVID-19), caused by severe acute respiratory syndrome coronavirus 2, during the pandemic altered the epidemiology of respiratory viruses. This study aimed to determine the changes in respiratory viruses among children hospitalized from 2018 to 2023. Nasopharyngeal specimens were collected from children aged under 15 years with fever and/or respiratory symptoms admitted to a medical institution in Fukushima Prefecture between January 2018 and December 2023. Eighteen respiratory viruses were detected using real-time reverse transcription-polymerase chain reaction. Overall, 1933 patients were included. Viruses were detected in 1377 (71.2%); of these, a single virus was detected in 906 (46.9%) and multiple viruses in 471 (24.3%). Among the viruses whose epidemics were temporarily suppressed, the epidemics of respiratory syncytial virus A and human parainfluenza virus type 3 (HPIV3) started earlier, and the epidemics of human metapneumovirus, HPIV1, and influenza A and C viruses resumed as behavioral restrictions for preventing COVID-19 eased. The median age of children with airway infection was significantly higher in the postpandemic group than in the prepandemic group (18.0 months vs. 21.0 months, p < 0.01). The median age of children infected with HPIV3 and human rhinovirus was significantly higher in the postpandemic group than in the prepandemic group. Strengthening of nonpharmaceutical interventions changed the epidemic dynamics of pediatric infectious diseases, with a trend toward older hospitalized children. Continuous monitoring of pediatric infectious disease outbreaks in hospitalized children can help prepare for the emergence of future viruses and pandemics.

The structural basis of protective and nonprotective human monoclonal antibodies targeting the parainfluenza virus type 3 hemagglutinin-neuraminidase

The structural basis of protective and nonprotective human monoclonal antibodies targeting the parainfluenza virus type 3 hemagglutinin-neuraminidase

Frequency of Bovine Respiratory Disease Complex Bacterial and Viral Agents Using Multiplex Real-Time qPCR in Quebec, Canada, from 2019 to 2023.

The bovine respiratory disease complex (BRD) is a multifactorial disease caused by various bacterial and viral pathogens. Using rapid pathogen detection techniques is helpful for tailoring therapeutic and preventive strategies in affected animals and herds. The objective of this study was to report the frequency of 10 pathogens by multiplex RT-qPCR on samples submitted for BRD diagnosis to a diagnostic laboratory (Biovet Inc., QC, Canada) in the Province of Quebec, Eastern Canada. From the 1st of January 2019 to the 31st of December 2023, a total of 1875 samples were analyzed. Most samples collected were individual samples (1547 of 1860 samples for which information was available (83.17%)), and the rest were from pooled samples of 2 (8.55%, n = 159) or ≥3 specimens (8.28%, n = 154). In 19.3% of the samples (n = 362), no pathogen was found, whereas 54.1% of samples had two or more different pathogens. Among the viruses, bovine coronavirus (BCV) was the most commonly found (27.5% of samples, n = 516), followed by bovine respiratory syncytial virus (BRSV) (17.7%, n = 332), whereas, for bacteria, Pasteurella multocida (50.1%, n = 940) and Mannheimia haemolytica (26.9%, n = 505) were the most common. The frequency of samples positive for Histophilus somni, Mycoplasmopsis bovis, influenza type D virus (IDV), bovine parainfluenza virus type 3 (BPI3V), bovine herpesvirus type 1 (BHV1), and bovine viral diarrhea virus (BVDV) was 22.6%, 22.4%, 4.6%, 4.3%, 2.7%, and 0.9%, respectively. In the multivariable Poisson regression model, the total number of pathogens increased with the number of animals in the pool, with an incidence risk ratio (IRR) of 1.15 (95% CI 1.02-1.29) and 1.32 (1.18-1.47) for 2 individuals in the pool and ≥3 individuals vs. individual samples, respectively. An increased number of pathogens were isolated in the winter season (IRR = 1.28 (95% CI 1.17-1.40)) compared to fall, and a lower number of pathogens were isolated in the summer compared to fall (IRR = 0.82 (95% CI 0.73-0.92)). These seasonal differences were mostly driven by the number of viruses isolated. This study gives interesting insights on the circulation of BRD pathogens in cattle from Eastern Canada.

Development of Primer Panels for Amplicon Sequencing of Human Parainfluenza Viruses Type 1 and 2.

Human parainfluenza viruses (hPIVs) are major contributors to respiratory tract infections in young children worldwide. Despite their global significance, genomic surveillance of hPIV1 and hPIV2 had not previously been conducted in Russia. This study aimed to develop a robust amplicon-based sequencing protocol for these viruses. The designed primer sets were tested on clinical samples containing hPIV RNA to evaluate their performance and efficiency. Sequencing results demonstrated high-quality genome data and efficient amplification across various Ct values. As a result, 41 hPIV1 and 13 hPIV2 near-complete genome sequences were successfully obtained from clinical specimens collected in Saint Petersburg (Russia). Phylogenetic analysis of the HN gene sequences showed that Russian hPIV1 strains clustered into clades II and III, while hPIV2 strains were distributed between clusters G1a and G3. The whole-genome-based trees confirmed the same distribution of the strains. These findings highlight the potential of our primer panels and contribute to a better understanding of the molecular characteristics and phylogenetic diversity of circulating hPIV strains. Notably, this study presents the first evolutionary analysis of hPIVs in Russia.

Coinfection of some respiratory viruses in cattle: An abattoir study.

To investigate the existence of bovine viral diarrhoea virus (BVDV), parainfluenza virus 3 (PIV-3) and respiratory syncytial virus (RSV) as well as its coinfections, a total of 420 pneumonic lung tissue samples were collected from slaughterhouses in three different areas. Samples were examined for the three viruses using antigen detection enzyme-linked immunosorbent assay (ELISA) test, and positive results were further confirmed using fluorescent antibody test and polymerase chain reaction. Prevalences detected were 10.47% for BVDV, 11.9% for parainfluenza virus type (PIV-3) and 7.6% for respiratory syncytial virus (RSV). Observed coinfections were bovine viral diarrhoea (BVD) and PIV-3 in 8.1%, BVD and RSV in only two samples (1%). Existence of all three viruses was detected in two samples (1%).Contribution:In this study, infection of the three viruses was confirmed in cattle and existence of its coinfection is documented for the first time.

Epidemiological, Clinical, and Genomic Traits of PIV in Hospitalized Children After the COVID-19 Pandemic in Wuhan, China.

Human parainfluenza virus (PIV) is a main cause of acute lower respiratory tract infections (ALRTIs), which contributes to childrens' mortality worldwide; however, the epidemiology of PIVs following the SARS-CoV-2 pandemic is still not clarified, and poses risks of potential outbreaks. Herein, we conducted a retrospective observational study from September 26, 2020 to September 30, 2023 to assess PIV epidemiology in Wuhan, China, as well as the clinical characteristics of PIV infections. In total, 14,065 inpatients with ALRTIs were enrolled, of which 936 were identified to have PIV infection. We also obtained 69 PIV3 RNA to reveal its molecular traits. An alteration in PIV season pattern away from spring and summer prevalence was noted, as well as a progressive rise in its detection rate. PIV-related ALRTIs were more prevalent in male patients. PIV3 was the dominant PIV type in recent years. In comparison with the phase before the cancellation of Dynamic Zero-COVID Policy in December 2022, symptoms after its repeal were milder. All Wuhan strains were classified with C3f lineage and possibly evolved from native strains in China. Additionally, some mutations, such as Q499P in protein hemagglutinin-neuraminidase, should be given further attention. In summary, our study demonstrates the clinical characteristics of PIVs and genomic traits of PIV3 in Wuhan, China, thus holds importance for the diagnosis and control of PIV infections in the post-pandemic era.

Exploring the host response in infected lung organoids using NanoString technology: A statistical analysis of gene expression data.

In this study, we used a three-dimensional airway "organ tissue equivalent" (OTE) model at an air-liquid interface (ALI) to mimic human airways. We investigated the effects of three viruses (Influenza A virus (IAV), Human metapneumovirus (MPV), and Parainfluenza virus type 3 (PIV3) on this model, incorporating various control conditions for data integrity. Our primary objective was to assess gene expression using the NanoString platform in OTE models infected with these viruses at 24- and 72-hour intervals, focusing on 773 specific genes. To enhance the comprehensiveness of our analysis, we introduced a novel algorithm, namely MAS (Magnitude-Altitude Score). This innovative approach uniquely combines biological significance, as indicated by fold changes in gene expression, with statistical rigor, as represented by adjusted p-values. By incorporating both dimensions, MAS ensures that the genes identified as differentially expressed are not mere statistical artifacts but hold genuine biological relevance, providing a more holistic understanding of the airway tissue response to viral infections. Our results unveiled distinct patterns of gene expression in response to viral infections. At 24 hours post-IAV infection, a robust interferon-stimulated gene (ISG) response was evident, marked by the upregulation of key genes including IFIT2, RSAD2, IFIT3, IFNL1, IFIT1, IFNB1, ISG15, OAS2, OASL, and MX1, collectively highlighting a formidable antiviral defense. MPV infection at the same time point displayed a dual innate and adaptive immune response, with highly expressed ISGs, immune cell recruitment signaled by CXCL10, and early adaptive immune engagement indicated by TXK and CD79A. In contrast, PIV3 infection at 24 hours triggered a transcriptional response dominated by ISGs, active immune cell recruitment through CXCL10, and inflammation modulation through OSM. The picture evolved at 72 hours post-infection. For IAV, ISGs and immune responses persisted, suggesting a sustained impact. MPV infection at this time point showed a shift towards IL17A and genes related to cellular signaling and immune responses, indicating adaptation to the viral challenge over time. In the case of PIV3, the transcriptional response remained interferon-centric, indicating a mature antiviral state. Our analysis underscored the pivotal role of ISGs across all infections and time points, emphasizing their universal significance in antiviral defense. Temporal shifts in gene expression indicative of adaptation and fine-tuning of the immune response. Additionally, the identification of shared and unique genes unveiled host-specific responses to specific pathogens. IAV exerted a sustained impact on genes from the initial 24 hours, while PIV3 displayed a delayed yet substantial genomic response, suggestive of a gradual and nuanced strategy.

Establishment of a Real-Time Reverse Transcription Recombinase-Aided Isothermal Amplification (qRT-RAA) Assay for the Rapid Detection of Bovine Respiratory Syncytial Virus.

Bovine respiratory syncytial virus (BRSV) is a significant cause of bovine respiratory disease, resulting in significant losses to the cattle industry. For rapid detection of BRSV, a real-time recombinase-aided isothermal amplification assay (qRT-RAA) based on the F gene of BRSV was developed in this study. The developed qRT-RAA assay showed good exponential amplification of the target fragment in 20 min at a constant temperature of 39 °C. And this assay displayed a high specificity for BRSV, without cross-reactions with Infectious Bovine Rhinotracheitis Virus (IBRV), Bovine Parainfluenza Virus Type 3 (BPIV3), Bovine Viral Diarrhea Virus (BVDV), and Bovine Coronavirus (BCoV). With the standard RNA of BRSV serving as a template, the limit of detection for qRT-RAA was 102 copies/μL. We examined ninety-seven clinical samples from cattle with respiratory disease using this method and determined a positive rate of 7.2% (7/97), consistent with results using the classical PCR method reported previously. A qRT-RAA assay for BRSV detection was established in this study. The method is specific and sensitive and can be completed within 20 min at 39 °C. These works demonstrate that the generated qRT-RAA assay is an effective diagnostic tool for rapidly detecting BRSV in resource-limited settings, which may be applied for the clinical detection of BRSV.

Virus Detection Using Functionalized Graphene Oxide Composites

In this study, we explore the optimization of Reduced Graphene Oxide (RGO)-based sensors for detecting respiratory viruses, employing an innovative thermal reduction process. Traditional methods of graphene oxide (GO) production often struggle with scalability and uniformity. Addressing these challenges, we utilize a common glass substrate within a controlled oven environment, achieving precise temperature regulation essential for consistent sensor properties. RGO produced at these specific conditions demonstrated significantly enhanced uniformity and electrical properties when compared to those produced under traditional, less controlled conditions. Expanding upon this base, we have now incorporated metals (such as Fe, Ag) into the graphene oxide matrix to create metal-graphene oxide composites. These composites were methodically synthesized and integrated into our existing Field-Effect Transistor (FET) sensors, allowing us to evaluate their enhanced responsiveness to Human Parainfluenza Virus types 2 and 3 (HPIV-2, HPIV-3), and Respiratory Syncytial Virus (RSV). Early experiments with these new composites show a marked improvement in sensor sensitivity and specificity, particularly against HPIV-3, suggesting a robust interaction between the virus and the sensor interface. The addition of metal composites not only fortifies the electrical and mechanical properties of the sensors but also significantly boosts their viral detection capabilities. This advancement confirms the reproducibility and reliability of the optimized RGO production process, reinforcing the importance of precise thermal control in producing not just stable and uniform RGO, but also highly effective biosensing platforms. These results pave the way for the potential mass production of RGO-based sensors, equipped with metal-graphene oxide composites, targeting high sensitivity and specificity in virus detection. This enhancement has profound implications for the broad applicability of RGO in diverse fields including electronics, chemical sensing, and gas sensing, contributing foundational knowledge for the advancement of graphene-based technologies. This research reaffirms the critical need for standardized production processes that ensure the consistency and efficiency of RGO and showcases the practical application of these processes in creating high-performance biosensors.

Derivatives of (–)-Cytisine with Thiourea Fragment. Synthesis and Antiviral Activity

New derivatives of the quinolizidine alkaloid, (–)-cytisine, with a substituted 2-pyridone ring and a thiourea moiety in the bispidin fragment of the molecule were synthesized. The ability of the synthesized cytisine-containing thioureas to inhibit the reproduction of human parainfluenza virus type 3 was assessed. It was found that the derivatives obtained by the reaction of benzoyl or phenyl isothiocyanate with (–)-cytisine, as well as its 9-bromo or 9,11-derivative, effectively suppress the reproduction of human parainfluenza virus type 3 (their selectivity indices are 56, 58 and 95, respectively), which confirms the promise of the chosen approach to synthetic modifications of the alkaloid (–)-cytisine in order to obtain effective antiviral agents on its basis.

Protective efficacy of a recombinant adenovirus expressing novel dual F and HN proteins of bovine parainfluenza virus type 3

Bovine parainfluenza virus type 3 (BPIV3) is a viral respiratory pathogen that infects cattle and causes significant economic losses. We generated a recombinant adenovirus called rHAd5-F + HN by expressing the fusion (F) and hemagglutinin-neuraminidase (HN) glycoprotein of BPIV3 using the human adenovirus serotype 5 (rHAd5). We evaluated its effects on humoral and cellular immune responses in mice (n = 45) and calves (n = 9). Serum antibody responses were assessed by enzyme-linked immunosorbent assay (ELISA), hemagglutination inhibition (HI), and neutralising antibodies (NAb). After boosting immunity with rHAd5-F + HN, mice produced significantly higher levels of antibodies against the BPIV3 genotype A and genotype C strains. The production of antibodies exceeded those produced by adenoviruses rHAd5-F and rHAd5-HN, which express the F and HN glycoprotein, respectively. The percentages of splenic CD3+/CD8+T lymphocytes and IL-4+ cytokines in rHAd5-F + HN mice were considerably higher than those in the control group. Mice immunised with rHAd5-F + HN exhibited much lower viral loads in the lungs and tracheas compared to the control group. Additionally, the lungs of mice vaccinated with rHAd5-F + HN showed no notable histopathological changes. On the other hand, rHAd5-F + HN produced a humoral immune response in calves. Following the booster intramuscular injection with the rHAd5-F + HN, the serum antibody levels against BPIV3 genotype C strain were 1:20 452, 1:1024, and 1:426 in calves, as detected by ELISA, HI, and NAb, respectively. The HI and NAb levels against the BPIV3 genotype A strain were 1:213 and 1:85 in calves, respectively. These results indicate that rHAd5-F + HN effectively induced immunity against BPIV3 infection.

Childhood Respiratory Illnesses Before and After COVID-19 Pandemic Restrictions.

Introduction With the COVID-19 pandemic, multiple studies described a significant drop in common respiratory viruses in children with the lockdown and restrictions. With the lifting of pandemic precautions, we had the ability to observe new patterns of respiratory illnesses in children and emergency department visits. Materials and methods We studied all respiratory nucleic acid amplification test results in emergency patients from a large metropolitan children's hospital from the years 2018 to 2023. The test included adenovirus, coronaviruses HKU1, NL63, 229E, OC43, metapneumovirus, rhinovirus or enterovirus, influenza A and B, parainfluenza virus types 1-4, respiratory syncytial virus, Bordetella pertussis, Bordetella parapertussis, Chlamydophila pneumoniae, and Mycoplasma pneumoniae. Coronavirus SARS CoV-2 became part of the respiratory panel in November of 2020. We reviewed pediatric emergency department census data to describe the trends before, during, and after COVID-19 pandemic restrictions. Results and conclusions Prior to 2020, there was a median of 1080 tests performed per week with an average positivity rate of 3.7-4.1%. During 2020, this dropped to 486 tests per week with a positivity rate of 1.74%. In 2021, after schools reopened, the median number of tests was 589 per week, with a positivity rate of 4.07%. After schools reopened without masks, the median tests per week were 817, with a positivity rate of 4.71%. Emergency department census data showed a large rebound in 2021 and 2022, with significantly earlier census peaks in these years. Common pediatric respiratory illnesses had an early seasonal spike in the years after the restrictions were lifted, with the most significant being the year in which local schools stopped wearing masks.

Next Generation RNA/Protein-Carrying Vector With Pleiotropic Activity.

Human parainfluenza virus type 2 (hPIV2), one of the causative agents of infantile common cold, is a non-segmented negative-sense RNA virus with a robust gene expression system. It infects recurrently throughout human life without causing severe disease. Because hPIV2 has a viral envelope that can carry ectopic proteins, we developed a non-propagative RNA/protein-carrying vector BC-PIV by deleting the F gene from hPIV2. BC-PIV can be vigorously proliferated in the stable packaging cell line Vero/BC-F cells expressing the hPIV2 F gene but not in other cells. BC-PIV can deliver exogenous gene(s) on a multigenic RNA genome as an inserted gene fragment(s) and simultaneously deliver exogenous protein(s) on its envelope in a membrane-anchored form. For example, influenza virus M2e protein, Ebola virus GP protein, and severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) spike protein were shown to be highly expressed in packaging cells and incorporated into the virion. The Ebola virus GP protein and SARS-CoV-2 spike protein, each delivered via BC-PIV, efficiently induced neutralising antibodies against each virus, even after prior treatment with recombinant BC-PIV in mice and hamsters, respectively. In this review, we describe the properties of BC-PIV as a promising vaccine vector, and also demonstrate its application as an anti-tumour virus.

Viral co-detection of influenza virus and other respiratory viruses in hospitalized Brazilian patients during the first three years of the coronavirus disease (COVID)-19 pandemic: an epidemiological profile.

In Brazil, few studies were performed regarding the co-detection of respiratory viruses in hospitalized patients. In this way, the study aimed to describe the epidemiological profile of hospitalized patients due to influenza virus infection that presented co-detection with another respiratory virus. The epidemiological analysis was made by collecting data from Open-Data-SUS. The study comprised patients infected by the influenza A or B virus with positive co-detection of another respiratory virus, such as adenovirus, bocavirus, metapneumovirus, parainfluenza virus (types 1, 2, 3, and 4), rhinovirus, and respiratory syncytial virus (RSV). The markers [gender, age, clinical signs and symptoms, comorbidities, need for intensive care unit (ICU) treatment, and need for ventilatory support] were associated with the chance of death. The data was collected during the first three years of the coronavirus disease (COVID)-19 pandemic-from December 19, 2019, to April 06, 2023. A total of 477 patients were included, among them, the influenza A virus was detected in 400 (83.9%) cases. The co-detection occurred, respectively, for RSV (53.0%), rhinovirus (14.0%), adenovirus (13.4%), parainfluenza virus type 1 (10.7%), parainfluenza virus type 3 (5.2%), metapneumovirus (3.8%), parainfluenza virus type 2 (3.6%), bocavirus (3.4%), and parainfluenza virus type 4 (1.5%). The co-detection rate was higher in the male sex (50.7%), age between 0-12 years of age (65.8%), and white individuals (61.8%). The most common clinical symptoms were cough (90.6%), dyspnea (78.8%), and fever (78.6%). A total of 167 (35.0%) people had at least one comorbidity, mainly cardiopathy (14.3%), asthma (8.4%), and diabetes mellitus (7.3%). The need for ICU treatment occurred in 147 (30.8%) cases, with most of them needing ventilatory support (66.8%), mainly non-invasive ones (57.2%). A total of 33 (6.9%) patients died and the main predictors of death were bocavirus infection (OR = 14.78 [95%CI = 2.84-76.98]), metapneumovirus infection (OR = 8.50 [95%CI = 1.86-38.78]), race (other races vs. white people) (OR = 3.67 [95%CI = 1.39-9.74]), cardiopathy (OR = 3.48 [95%CI = 1.13-10.71]), and need for ICU treatment (OR = 7.64 [95%CI = 2.44-23.92]). Co-detection between the influenza virus and other respiratory viruses occurred, mainly with RSV, rhinovirus, and adenovirus being more common in men, white people, and in the juvenile phase. Co-detection of influenza virus with bocavirus and metapneumovirus was associated with an increased chance of death. Other factors such as race, cardiopathy, and the need for an ICU were also associated with a higher chance of death.