bluuebunny/crossref_metadata_2025 huggingface dataset:
Effects of Harmful Algal Blooms on Fish and Shellfish Species: A Case Study of New Zealand in a Changing Environment
Authors: Rolton Anne , Rhodes Lesley , Hutson Kate S. , Biessy Laura , Bui Tony , MacKenzie Lincoln , Symonds Jane E. , Smith Kirsty F.
Published: May 2022

Harmful algal blooms (HABs) have wide-ranging environmental impacts, including on aquatic species of social and commercial importance. In New Zealand (NZ), strategic growth of the aquaculture industry could be adversely affected by the occurrence of HABs. This review examines HAB species which are known to bloom both globally and in NZ and their effects on commercially important shellfish and fish species. Blooms of Karenia spp. have frequently been associated with mortalities of both fish and shellfish in NZ and the sub-lethal effects of other genera, notably Alexandrium spp., on shellfish (which includes paralysis, a lack of byssus production, and reduced growth) are also of concern. Climate change and anthropogenic impacts may alter HAB population structure and dynamics, as well as the physiological responses of fish and shellfish, potentially further compromising aquatic species. Those HAB species which have been detected in NZ and have the potential to bloom and harm marine life in the future are also discussed. The use of environmental DNA (eDNA) and relevant bioassays are practical tools which enable early detection of novel, problem HAB species and rapid toxin/HAB screening, and new data from HAB monitoring of aquaculture production sites using eDNA are presented. As aquaculture grows to supply a sizable proportion of the world’s protein, the effects of HABs in reducing productivity is of increasing significance. Research into the multiple stressor effects of climate change and HABs on cultured species and using local, recent, HAB strains is needed to accurately assess effects and inform stock management strategies.
Novel Anatomical Guidelines on Botulinum Neurotoxin Injection for Wrinkles in the Nose Region
Authors: Yi Kyu-Ho , Lee Ji-Hyun , Hu Hye-Won , Kim Hee-Jin
Published: May 2022

Botulinum neurotoxin injection surrounding the nose area is frequently used in aesthetic settings. However, there is a shortage of thorough anatomical understanding that makes it difficult to treat wrinkles in the nose area. In this study, the anatomical aspects concerning the injection of botulinum neurotoxin into the nasalis, procerus, and levator labii superioris alaeque muscles are assessed. In addition, the present knowledge on localizing the botulinum neurotoxin injection point from a newer anatomy study is assessed. It was observed that, for the line-associated muscles in the nose region, the injection point may be more precisely defined. The optimal injection sites are the nasalis, procerus, and levator labii superioris alaeque muscles, and the injection technique is advised. We advise the best possible injection sites in association with anatomical standards for commonly injected muscles to increase efficiency in the nose region by removing the wrinkles. Similarly, these suggestions support a more precise procedure.
The Utility of the Shock Index for Predicting Survival, Function and Health Status Outcomes in Major Trauma Patients: A Registry-Based Cohort Study
Authors: Wikström Lena , Kander Thomas , Gabbe Belinda J.
Published: May 2022

The shock index (SI; heart rate/systolic blood pressure) has been found to predict mortality in trauma patients. The aims of this study were to establish whether the SI improved the prediction of an existing model for both mortality and longer-term outcomes in major trauma patients. In total, 29,574 adult (>15 years) major trauma patients were included from the Victorian State Trauma Registry with a date of injury from July 2009 to June 2019. Outcomes of interest were survival to hospital discharge and function and health status at 6 months post-injury. Survival and function were analysed using measures of discrimination and calibration, whereas health status was assessed with R2 and MRSE measures. The area under the receiver operating characteristic curve (AUC) of the VSTR survival model improved when the SI was added (AUC 0.797 (0.787–0.807) versus AUC 0.807 (0.797–0.816), p < 0.001). For the prediction of functional outcome 6 months post-injury, the inclusion of the SI increased the AUC marginally (AUC 0.795 (0.786–0.803) versus AUC 0.799 (0.791–0.807), p < 0.001). When predicting in-hospital mortality and health status 6 months post-injury, including the SI resulted in a slightly better fit to the registry-risk adjustment model. In the future, external validation and the exploration of other models to improve prediction outcomes are warranted.
Systematic Review and Meta-Analysis on the Infection Rates of Schistosome Transmitting Snails in Southern Africa
Authors: Nwoko Onyekachi Esther , Kalinda Chester , Chimbari Moses John
Published: May 2022

Efforts to interrupt and eliminate schistosomiasis as a public health problem have increased in several Southern African countries. A systematic review was carried out on the infection rates of snails that cause schistosomiasis in humans. The searches were conducted in PubMed, Web of Science, and Scopus databases, using the PRISMA guidelines from inception to 24 February 2022. The study quality was assessed by using the Joanna Briggs Institute prevalence critical appraisal checklist. Pooled infection rates were estimated by using an inverse variance heterogeneity model, while heterogeneity was determined by using Cochran’s Q test and Higgins i2 statistics. A total of 572 articles were screened, but only 28 studies were eligible for inclusion based on predetermined criteria. In the selected studies, 82,471 Bulinus spp. and 16,784 Biomphalaria spp. snails were screened for cercariae. The pooled infectivity of schistosome intermediate host snails, Biomphalaria spp., and Bulinus spp. were 1%, 2%, and 1%, respectively. Snail infection rates were higher in the 1900s compared to the 2000s. A Luis Furuya–Kanamori index of 3.16 indicated publication bias, and a high level of heterogeneity was observed. Although snail infectivity in Southern Africa is relatively low, it falls within the interval of common snail infection rates, thus indicating the need for suitable snail control programs that could interrupt transmission and achieve elimination.
Needs Assessment of Southeastern United States Vector Control Agencies: Capacity Improvement Is Greatly Needed to Prevent the Next Vector-Borne Disease Outbreak
Authors: Dye-Braumuller Kyndall C. , Gordon Jennifer R. , Johnson Danielle , Morrissey Josie , McCoy Kaci , Dinglasan Rhoel R. , Nolan Melissa S.
Published: May 2022

A national 2017 vector control capacity survey was conducted to assess the United States’ (U.S.’s) ability to prevent emerging vector-borne disease. Since that survey, the southeastern U.S. has experienced continued autochthonous exotic vector-borne disease transmission and establishment of invasive vector species. To understand the current gaps in control programs and establish a baseline to evaluate future vector control efforts for this vulnerable region, a focused needs assessment survey was conducted in early 2020. The southeastern U.S. region was targeted, as this region has a high probability of novel vector-borne disease introduction. Paper copies delivered in handwritten envelopes and electronic copies of the survey were delivered to 386 unique contacts, and 150 returned surveys were received, corresponding to a 39% response rate. Overall, the survey found vector control programs serving areas with over 100,000 residents and those affiliated with public health departments had more core capabilities compared to smaller programs and those not affiliated with public health departments. Furthermore, the majority of vector control programs in this region do not routinely monitor for pesticide resistance. Taken as a whole, these results suggest that the majority of the southeastern U.S. is vulnerable to vector-borne disease outbreaks. Results from this survey raise attention to the critical need of providing increased resources to bring all vector control programs to a competent level, ensuring that public health is protected from the threat of vector-borne disease.
Extended Gravity Constraints at Different Scales
Authors: Alexeyev Stanislav , Prokopov Vyacheslav
Published: May 2022

We review a set of the possible ways to constrain extended gravity models at Galaxy clusters scales (the regime of dark energy explanations and comparison with ΛCDM), for black hole shadows, gravitational wave astronomy, binary pulsars, the Solar system and a Large Hadron Collider (consequences for high-energy physics at TeV scale). The key idea is that modern experimental and observational precise data provide us with the chance to go beyond general relativity.
Pull-Through Ureteroneocystostomy for Very Small En Bloc Kidney Transplants from Donors Weighing ≤ 5 kg
Authors: Nghiem Dai D.
Published: May 2022

Background: Urologic complications are the most dreaded complications of renal transplantation, particularly when pediatric en bloc kidneys (EBKs) are used. Current techniques of ureteroneocystostomy (UNC) are not applicable to the very small ureters of very small en bloc kidneys. We reviewed our experience with the pull-through ureteroneocystostomy in kidney transplantation from donors under or equal to 5 kg weight. Material and Methods: The technique was used in 32 EBKs. Complications and 4-year graft survival are discussed. Results: One single graft thrombosed and required nephrectomy. The remaining kidneys provided good renal function. Hematuria was transient in five patients and did not require fulguration. No leakage was experienced. Delayed graft function occurred in 16% of cases. No primary nonfunction was noted. During the follow-up period, hydronephrosis and/or pyelonephritis were not observed. The 4-year graft survival was 95% with serum creatinine levels averaging 0.9 mg/dl. Conclusions: The procedure proved to be safe and reproducible. It can be applied to the ureteral re-implantation of very small EBKs.
Efficacy of the Sentinox Spray in Reducing Viral Load in Mild COVID-19 and Its Virucidal Activity against Other Respiratory Viruses: Results of a Randomized Controlled Trial and an In Vitro Study
Authors: Panatto Donatella , Orsi Andrea , Bruzzone Bianca , Ricucci Valentina , Fedele Guido , Reiner Giorgio , Giarratana Nadia , Domnich Alexander , Icardi Giancarlo ,   STX Study Group
Published: May 2022

Sentinox (STX) is an acid-oxidizing solution containing hypochlorous acid in spray whose virucidal activity against SARS-CoV-2 has been demonstrated. In this paper, results of a randomized controlled trial (RCT) on the efficacy of STX in reducing viral load in mild COVID-19 patients (NCT04909996) and a complementary in vitro study on its activity against different respiratory viruses are reported. In the RCT, 57 patients were randomized (1:1:1) to receive STX three (STX-3) or five (STX-5) times/day plus standard therapy or standard therapy only (controls). Compared with controls, the log10 load reduction in groups STX-3 and STX-5 was 1.02 (p = 0.14) and 0.18 (p = 0.80), respectively. These results were likely driven by outliers with extreme baseline viral loads. When considering subjects with baseline cycle threshold values of 20–30, STX-3 showed a significant (p = 0.016) 2.01 log10 reduction. The proportion of subjects that turned negative by the end of treatment (day 5) was significantly higher in the STX-3 group than in controls, suggesting a shorter virus clearance time. STX was safe and well-tolerated. In the in vitro study, ≥99.9% reduction in titers against common respiratory viruses was observed. STX is a safe device with large virucidal spectrum and may reduce viral loads in mild COVID-19 patients.
Bluetongue Virus Infection of Goats: Re-Emerged European Serotype 8 vs. Two Atypical Serotypes
Authors: Ries Christina , Beer Martin , Hoffmann Bernd
Published: May 2022

In recent years, numerous atypical Bluetongue virus (BTV) strains have been discovered all around the world. Atypical BTV strains are phylogenetically distinct from the classical BTV serotypes 1–24 and differ in terms of several biological features. For the first time, the atypical strains BTV-25-GER2018 and BTV-33-MNG3/2016 as well as the re-emerged classical strain BTV-8-GER2018 were evaluated comparatively in a pathogenesis study in goats—the natural host of atypical BTV. A substantial number of in-contact animals were included in this study to detect potential contact transmissions of the virus. After infection, EDTA blood, ocular, nasal and oral swab samples as well as serum were collected regularly and were used for virological and serological analyses, respectively. Our study showed differences in the immunological reaction between the two atypical BTV strains (no group-specific antibody detection) and the classical BTV strain BTV-8-GER2018 (group-specific antibody detection). Furthermore, we observed an increase in the total WBC count (neutrophils and lymphocytes) in goats infected with the atypical BTV strains. No horizontal transmission was seen for all three strains. Our study suggests that the atypical BTVs used in the trial differ from classical BTVs in their immunopathogenesis. However, no evidence of direct contact transmission was found.
Impaired Vagal Activity in Long-COVID-19 Patients
Authors: Acanfora Domenico , Nolano Maria , Acanfora Chiara , Colella Camillo , Provitera Vincenzo , Caporaso Giuseppe , Rodolico Gabriele Rosario , Bortone Alessandro Santo , Galasso Gennaro , Casucci Gerardo
Published: May 2022

Long-COVID-19 refers to the signs and symptoms that continue or develop after the “acute COVID-19” phase. These patients have an increased risk of multiorgan dysfunction, readmission, and mortality. In Long-COVID-19 patients, it is possible to detect a persistent increase in D-Dimer, NT-ProBNP, and autonomic nervous system dysfunction. To verify the dysautonomia hypothesis in Long-COVID-19 patients, we studied heart rate variability using 12-lead 24-h ECG monitoring in 30 Long-COVID-19 patients and 20 No-COVID patients. Power spectral analysis of heart rate variability was lower in Long-COVID-19 patients both for total power (7.46 ± 0.5 vs. 8.08 ± 0.6; p < 0.0001; Cohens-d = 1.12) and for the VLF (6.84 ± 0.8 vs. 7.66 ± 0.6; p < 0.0001; Cohens-d = 1.16) and HF (4.65 ± 0.9 vs. 5.33 ± 0.9; p = 0.015; Cohens-d = 0.76) components. The LF/HF ratio was significantly higher in Long-COVID-19 patients (1.46 ± 0.27 vs. 1.23 ± 0.13; p = 0.001; Cohens-d = 1.09). On multivariable analysis, Long-COVID-19 is significantly correlated with D-dimer (standardized β-coefficient = 0.259), NT-ProBNP (standardized β-coefficient = 0.281), HF component of spectral analysis (standardized β-coefficient = 0.696), and LF/HF ratio (standardized β-coefficient = 0.820). Dysautonomia may explain the persistent symptoms in Long COVID-19 patients. The persistence of a procoagulative state and an elevated myocardial strain could explain vagal impairment in these patients. In Long-COVID-19 patients, impaired vagal activity, persistent increases of NT-ProBNP, and a prothrombotic state require careful monitoring and appropriate intervention.
Comparing Influenza Virus Biology for Understanding Influenza D Virus
Authors: Skelton Raegan M. , Huber Victor C.
Published: May 2022

The newest type of influenza virus, influenza D virus (IDV), was isolated in 2011. IDV circulates in several animal species worldwide, causing mild respiratory illness in its natural hosts. Importantly, IDV does not cause clinical disease in humans and does not spread easily from person to person. Here, we review what is known about the host–pathogen interactions that may limit IDV illness. We focus on early immune interactions between the virus and infected host cells in our summary of what is known about IDV pathogenesis. This work establishes a foundation for future research into IDV infection and immunity in mammalian hosts.
SARS-CoV-2 Infection and Possible Neonatal Neurological Outcomes: A Literature Review
Authors: de Moraes Flávia Maciel , de Souza Julia Werneck Paulino Soares , Alves Letícia Pires , de Siqueira Milena Ferreira Ribeiro , dos Santos Ana Paula Aguiar , de Carvalho Berardo Mariana Monteiro , Granja Marcelo Gomes , de Castro-Faria-Neto Hugo Caire
Published: May 2022

The virus responsible for COVID-19 is designated “severe acute respiratory syndrome coronavirus 2” (SARS-CoV-2), a highly transmissible and pathogenic coronavirus. Although people of all ages are susceptible to SARS-CoV-2 infection, clinical manifestations may vary with age. The response of neonates to SARS-CoV-2 infection or exposure differs from that of children and adults. Encephalitis due to viral infections in the central nervous system (CNS) and childhood multisystem inflammatory syndrome (MIS-C) are some of the possible neonatal consequences of SARS-CoV-2 infection. This review aims to verify possible neonatal neurological outcomes after SARS-CoV-2 infection. Overall, the cellular and molecular basis of the neurological sequelae of SARS-CoV-2 in neonates remains unclear, and attempts to elucidate the pathophysiology of COVID-19 involve a comparison with the mechanism of other viral diseases. There are a considerable number of case reports in the literature exploring neurological outcomes in the neonatal period. In this review, we present possible effects of SARS-CoV-2 in neonates, emphasizing the importance of monitoring this group. The mechanisms of SARS-CoV-2 entry into the CNS have not yet been fully elucidated, and the potential severity of SARS-CoV-2 infection in neonates, as well as the possible short- and long-term neurological sequelae, remain unclear.
Robust AAV Genotyping Based on Genetic Distances in Rep Gene That Are Maintained by Ubiquitous Recombination
Authors: Beloukhova Marina I. , Lukashev Alexander N. , Volchkov Pavel Y. , Zamyatnin Andrey A. , Deviatkin Andrei A.
Published: May 2022

Adeno-associated viruses (AAVs) are a convenient tool for gene therapy delivery. According to the current classification, they are divided into the species AAV A and AAV B within the genus Dependoparvovirus. Historically AAVs were also subdivided on the intraspecies level into 13 serotypes, which differ in tissue tropism and targeted gene delivery capacity. Serotype, however, is not a universal taxonomic category, and their assignment is not always robust. Cross-reactivity has been shown, indicating that classification could not rely on the results of serological tests alone. Moreover, since the isolation of AAV4, all subsequent AAVs were subdivided into serotypes based primarily on genetic differences and phylogenetic reconstructions. An increased interest in the use of AAV as a gene delivery tool justifies the need to improve the existing classification. Here, we suggest genotype-based AAV classification below the species level based on the rep gene. A robust threshold was established as 10% nt differences within the 1248 nt genome fragment, with 4 distinct AAV genotypes identified. This distinct sub-species structure is maintained by ubiquitous recombination within, but not between, rep genes of the suggested genotypes.
Signaling Pathway Reporter Screen with SARS-CoV-2 Proteins Identifies nsp5 as a Repressor of p53 Activity
Authors: Kumar Abhishek , Grams Tristan R. , Bloom David C. , Toth Zsolt
Published: May 2022

The dysregulation of host signaling pathways plays a critical role in severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection and viral pathogenesis. While a number of viral proteins that can block type I IFN signaling have been identified, a comprehensive analysis of SARS-CoV-2 proteins in the regulation of other signaling pathways that can be critical for viral infection and its pathophysiology is still lacking. Here, we screened the effect of 21 SARS-CoV-2 proteins on 10 different host signaling pathways, namely, Wnt, p53, TGFβ, c-Myc, Hypoxia, Hippo, AP-1, Notch, Oct4/Sox2, and NF-κB, using a luciferase reporter assay. As a result, we identified several SARS-CoV-2 proteins that could act as activators or inhibitors for distinct signaling pathways in the context of overexpression in HEK293T cells. We also provided evidence for p53 being an intrinsic host restriction factor of SARS-CoV-2. We found that the overexpression of p53 is capable of reducing virus production, while the main viral protease nsp5 can repress the transcriptional activity of p53, which depends on the protease function of nsp5. Taken together, our results provide a foundation for future studies, which can explore how the dysregulation of specific signaling pathways by SARS-CoV-2 proteins can control viral infection and pathogenesis.
Analysis of Human Papillomavirus (HPV) and Polyomaviruses (HPyVs) in Adenoid Cystic Carcinoma (AdCC) of the Head and Neck Region Reveals Three HPV-Positive Cases with Adenoid Cystic-like Features
Authors: Zupancic Mark , Holzhauser Stefan , Cheng Liquin , Ramqvist Torbjörn , Du Juan , Friesland Signe , Näsman Anders , Dalianis Tina
Published: May 2022

An aetiological role of human papillomavirus (HPV) and/or human polyomaviruses (HPyVs) has been proposed in adenoid cystic carcinoma (AdCC). Moreover, HPV-related multiphenotypic carcinoma (HMSC) was recently introduced as an emerging entity of the sinonasal region. Here, we primarily want to study the role of HPV/HPyV in a large AdCC cohort and, secondly, possibly identify and characterize HMSC. Tumour DNA from 68 patients initially diagnosed with AdCC between 2000 and 2012 was, therefore, tested for 27 HPV types and 10 HPyVs. HPV DNA-positive samples were micromorphologically re-evaluated, further stained for p16INK4a, S100, p63 and CD117 and tested for the presence of the MYB-NFIB fusion transcript. Notably, no samples were HPyV-positive, while one sinonasal and two tonsillar carcinomas were HPV- and p16-positive. After re-evaluating the micromorphology, immunohistochemistry and presence of fusion transcripts, all tumours had the same appearance and fitted within the diagnosis of HMSC, but in all these three cases, the morphology of the HMSC and basaloid squamous cell carcinoma was overlapping. We conclude that HPV and HPyV have no major role in AdCC. However, based on our data, we also suggest that HMSC should be considered as a basaloid variant of squamous cell carcinoma, and not its own entity, until better characterized.
Jingmen Tick Virus in Ticks from Kenya
Authors: Ogola Edwin O. , Kopp Anne , Bastos Armanda D. S. , Slothouwer Inga , Marklewitz Marco , Omoga Dorcus , Rotich Gilbert , Getugi Caroline , Sang Rosemary , Torto Baldwyn , Junglen Sandra , Tchouassi David P.
Published: May 2022

Jingmen tick virus (JMTV) is an arbovirus with a multisegmented genome related to those of unsegmented flaviviruses. The virus first described in Rhipicephalus microplus ticks collected in Jingmen city (Hubei Province, China) in 2010 is associated with febrile illness in humans. Since then, the geographic range has expanded to include Trinidad and Tobago, Brazil, and Uganda. However, the ecology of JMTV remains poorly described in Africa. We screened adult ticks (n = 4550, 718 pools) for JMTV infection by reverse transcription polymerase chain reaction (RT-PCR). Ticks were collected from cattle (n = 859, 18.88%), goats (n = 2070, 45.49%), sheep (n = 1574, 34.59%), and free-ranging tortoises (Leopard tortoise, Stigmochelys pardalis) (n = 47, 1.03%) in two Kenyan pastoralist-dominated areas (Baringo and Kajiado counties) with a history of undiagnosed febrile human illness. Surprisingly, ticks collected from goats (0.3%, 95% confidence interval (CI) 0.1–0.5), sheep (1.8%, 95% CI 1.2–2.5), and tortoise (74.5%, 95% CI 60.9–85.4, were found infected with JMTV, but ticks collected from cattle were all negative. JMTV ribonucleic acid (RNA) was also detected in blood from tortoises (66.7%, 95% CI 16.1–97.7). Intragenetic distance of JMTV sequences originating from tortoise-associated ticks was greater than that of sheep-associated ticks. Phylogenetic analyses of seven complete-coding genome sequences generated from tortoise-associated ticks formed a monophyletic clade within JMTV strains from other countries. In summary, our findings confirm the circulation of JMTV in ticks in Kenya. Further epidemiological surveys are needed to assess the potential public health impact of JMTV in Kenya.
Clearance of Persistent SARS-CoV-2 RNA Detection in a NFκB-Deficient Patient in Association with the Ingestion of Human Breast Milk: A Case Report
Authors: Sabino Janine S. , Amorim Mariene R. , de Souza William M. , Marega Lia F. , Mofatto Luciana S. , Toledo-Teixeira Daniel A. , Forato Julia , Stabeli Rodrigo G. , Costa Maria Laura , Spilki Fernando R. , Sabino Ester C. , Faria Nuno R. , Benites Bruno D. , Addas-Carvalho Marcelo , Stucchi Raquel S. B. , Vasconcelos Dewton M. , Weaver Scott C. , Granja Fabiana , Proenca-Modena José Luiz , Vilela Maria Marluce dos S.
Published: May 2022

Currently, there are no evidence-based treatment options for long COVID-19, and it is known that SARS-CoV-2 can persist in part of the infected patients, especially those with immunosuppression. Since there is a robust secretion of SARS-CoV-2-specific highly-neutralizing IgA antibodies in breast milk, and because this immunoglobulin plays an essential role against respiratory virus infection in mucosa cells, being, in addition, more potent in neutralizing SARS-CoV-2 than IgG, here we report the clinical course of an NFκB-deficient patient chronically infected with the SARS-CoV-2 Gamma variant, who, after a non-full effective treatment with plasma infusion, received breast milk from a vaccinated mother by oral route as treatment for COVID-19. After such treatment, the symptoms improved, and the patient was systematically tested negative for SARS-CoV-2. Thus, we hypothesize that IgA and IgG secreted antibodies present in breast milk could be useful to treat persistent SARS-CoV-2 infection in immunodeficient patients.
Assessment of Life Cycle Modeling Systems as Prediction Tools for a Possible Attenuation of Recombinant Ebola Viruses
Authors: Bodmer Bianca S. , Hoenen Thomas
Published: May 2022

Ebola virus (EBOV) causes hemorrhagic fever in humans with high case fatality rates. In the past, a number of recombinant EBOVs expressing different reporters from additional transcription units or as fusion proteins have been rescued. These viruses are important tools for the study of EBOV, and their uses include high throughput screening approaches, the analysis of intercellular localization of viral proteins and of tissue distribution of viruses, and the study of pathogenesis in vivo. However, they all show, at least in vivo, attenuation compared to wild type virus, and the basis of this attenuation is only poorly understood. Unfortunately, rescue of these viruses is a lengthy and not always successful process, and working with them is restricted to biosafety level (BSL)-4 laboratories, so that the search for non-attenuated reporter-expressing EBOVs remains challenging. However, several life cycle modeling systems have been developed to mimic different aspects of the filovirus life cycle under BSL-1 or -2 conditions, but it remains unclear whether these systems can be used to predict the viability and possible attenuation of recombinant EBOVs. To address this question, we systematically fused N- or C-terminally either a flag-HA tag or a green fluorescent protein (GFP) to different EBOV proteins, and analyzed the impact of these additions with respect to protein function in life cycle modeling systems. Based on these results, selected recombinant EBOVs encoding these tags/proteins were then rescued and characterized for a possible attenuation in vitro, and results compared with data from the life cycle modeling systems. While the results for the small molecular tags showed mostly good concordance, GFP-expressing viruses were more attenuated than expected based on the results from the life cycle modeling system, demonstrating a limitation of these systems and emphasizing the importance of work with infectious virus. Nevertheless, life cycle modeling system remain useful tools to exclude non-viable tagging strategies.
Infectious Bronchitis Virus Nsp14 Degrades JAK1 to Inhibit the JAK-STAT Signaling Pathway in HD11 Cells
Authors: Ma Peng , Gu Kui , Li Hao , Zhao Yu , Li Chao , Wen Renqiao , Zhou Changyu , Lei Changwei , Yang Xin , Wang Hongning
Published: May 2022

Coronaviruses (CoVs) are RNA viruses that can infect a wide range of animals, including humans, and cause severe respiratory and gastrointestinal disease. The Gammacoronavirus avian infectious bronchitis virus (IBV) causes acute and contagious diseases in chickens, leading to severe economic losses. Nonstructural protein 14 (Nsp14) is a nonstructural protein encoded by the CoV genome. This protein has a regulatory role in viral virulence and replication. However, the function and mechanism of IBV Nsp14 in regulating the host’s innate immune response remain unclear. Here we report that IBV Nsp14 was a JAK-STAT signaling pathway antagonist in chicken macrophage (HD11) cells. In these cells, Nsp14 protein overexpression blocked IBV suppression induced by exogenous chIFN-γ treatment. Meanwhile, Nsp14 remarkably reduced interferon-gamma-activated sequence (GAS) promoter activation and chIFN-γ-induced interferon-stimulated gene expression. Nsp14 impaired the nuclear translocation of chSTAT1. Furthermore, Nsp14 interacted with Janus kinase 1 (JAK1) to degrade JAK1 via the autophagy pathway, thereby preventing the activation of the JAK-STAT signaling pathway and facilitating viral replication. These results indicated a novel mechanism by which IBV inhibits the host antiviral response and provide new insights into the selection of antiviral targets against CoV.
HSV-1 Infection of Epithelial Dendritic Cells Is a Critical Strategy for Interfering with Antiviral Immunity
Authors: Gao Yang , Cheng Jishuai , Xu Xingli , Li Xueqi , Zhang Jingjing , Ma Danjing , Jiang Guorun , Liao Yun , Fan Shengtao , Niu Zhenye , Yue Rong , Chang Penglan , Zeng Fengyuan , Duan Suqin , Meng Ziyan , Xu Xiangxiong , Li Xinghang , Li Dandan , Yu Li , Ping Lifen , Zhao Heng , Guo Mingtian , Wang Lichun , Wang Yafang , Zhang Ying , Li Qihan
Published: May 2022

Herpes simplex virus type 1 (HSV-1), an α subgroup member of the human herpesvirus family, infects cells via the binding of its various envelope glycoproteins to cellular membrane receptors, one of which is herpes virus entry mediator (HVEM), expressed on dendritic cells. Here, HVEM gene-deficient mice were used to investigate the immunologic effect elicited by the HSV-1 infection of dendritic cells. Dendritic cells expressing the surface marker CD11c showed an abnormal biological phenotype, including the altered transcription of various immune signaling molecules and inflammatory factors associated with innate immunity after viral replication. Furthermore, the viral infection of dendritic cells interfered with dendritic cell function in the lymph nodes, where these cells normally play roles in activating the T-cell response. Additionally, the mild clinicopathological manifestations observed during the acute phase of HSV-1 infection were associated with viral replication in dendritic cells.
SARS-CoV-2 Seroconversion in an Adult Horse with Direct Contact to a COVID-19 Individual
Authors: Pusterla Nicola , Chaillon Antoine , Ignacio Caroline , Smith Davey M. , Barnum Samantha , Lawton Kaila O. Y. , Smith Greg , Pickering Bradley
Published: May 2022

The authors report on a possible direct exposure to SARS-CoV-2 from a COVID-19-positive individual to an adult horse. The individual, diagnosed with COVID-19 (Delta B.1.617.2), had daily contact to her two horses prior to and during the development of clinical disease. None of the two horses developed abnormal clinical signs or had detectable SARS-CoV-2 in blood, nasal secretion, or feces via RT-qPCR. However, one of the two horses showed close temporal seroconversion to SARS-CoV-2 using a protein-based ELISA and the plaque reduction neutralization test. The results suggest that horses can become silently infected with SARS-CoV-2 following close contact with humans infected with SARS-CoV-2. As a precautionary measure, humans infected with SARS-CoV-2 should avoid close contact with equids and other companion animals during the time of their illness to prevent viral transmission.
Development of DNA Vaccine Candidate against SARS-CoV-2
Authors: Wang Xingyun , Rcheulishvili Nino , Cai Jie , Liu Cong , Xie Fengfei , Hu Xing , Yang Nuo , Hou Mengqi , Papukashvili Dimitri , He Yunjiao , Wang Peng George
Published: May 2022

Despite the existence of various types of vaccines and the involvement of the world’s leading pharmaceutical companies, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) remains the most challenging health threat in this century. Along with the increased transmissibility, new strains continue to emerge leading to the need for more vaccines that would elicit protectiveness and safety against the new strains of the virus. Nucleic acid vaccines seem to be the most effective approach in case of a sudden outbreak of infection or the emergence of a new strain as it requires less time than any conventional vaccine development. Hence, in the current study, a DNA vaccine encoding the trimeric prefusion-stabilized ectodomain (S1+S2) of SARS-CoV-2 S-protein was designed by introducing six additional prolines mutation, termed HexaPro. The three-dose regimen of designed DNA vaccine immunization in mice demonstrated the generation of protective antibodies.
Porcine Epidemic Diarrhea Virus Infection Induces Autophagosome Formation but Inhibits Autolysosome Formation during Replication
Authors: Park Jae-Yeon , Ryu Jihoon , Hong Eui-Ju , Shin Hyun-Jin
Published: May 2022

In this study, we investigated the correlation between the mechanism involved in porcine epidemic diarrhea virus (PEDV) replication and autophagic flux. In this study, we found that as PEDV replicated, production of LC3-II was significantly induced up to 24 h post-infection (hpi). Interestingly, although there was significant production of LC3-II, greater p62 accumulation was simultaneously found. Pretreatment with rapamycin significantly induced PEDV replication, but autolysosome formation was reduced. These results were confirmed by the evaluation of ATG5/ATG12 and LAMP1/LAMP2. Taken together, we conclude that PEDV infection induces autophagosome formation but inhibits autolysosome formation during replication.
Molecular Analysis of Caprine Enterovirus Circulating in China during 2016–2021: Evolutionary Significance
Authors: Chang Xiaoran , Lin Qian , Zhang Qun , Hu Junying , Tursun Gulbahar , Deng Yingrui , Guo Chunguang , Wang Xinping
Published: May 2022

Here, we report the characterization of 13 novel caprine/ovine enterovirus strains isolated from different regions in China during 2016–2021. Immunoperoxidase monolayer assay showed that these viral strains shared strong cross-reaction with the previously reported caprine enterovirus CEV-JL14. Alignment analysis of the complete nucleotide sequences revealed 79.2%–87.8% and 75.0%–76.7% sequence identity of these novel caprine enterovirus strains to CEV-JL14 and TB4-OEV, respectively. Phylogenetic analyses clustered these novel strains to EV-G based on the amino acid sequences of P1 and 2C+3CD. Moreover, phylogenetic analysis of these caprine enterovirus strains identified three new EV-G types using VP1 sequences. These results demonstrate the genetic variations and the evolution of caprine enterovirus.
Inactivation Methods for Experimental Nipah Virus Infection
Authors: Widerspick Lina , Vázquez Cecilia Alejandra , Niemetz Linda , Heung Michelle , Olal Catherine , Bencsik András , Henkel Christoph , Pfister Anneke , Brunetti Jesús Emanuel , Kucinskaite-Kodze Indre , Lawrence Philip , Muñoz Fontela César , Diederich Sandra , Escudero-Pérez Beatriz
Published: May 2022

Nipah virus (NiV) is an emerging zoonotic paramyxovirus that causes severe disease in humans and livestock. Due to its high pathogenicity in humans and the lack of available vaccines and therapeutics, NiV needs to be handled in biosafety level 4 (BSL-4) laboratories. Safe inactivation of samples containing NiV is thus necessary to allow further processing in lower containment areas. To date, there is only limited information available on NiV inactivation methods validated by BSL-4 facilities that can be used as a reference. Here, we compare some of the most common inactivation methods in order to evaluate their efficacy at inactivating NiV in infected cells, supernatants and organs. Thus, several physical and chemical inactivation methods, and combinations thereof, were assessed. Viral replication was monitored for 3 weeks and NiV presence was assessed by RT-qPCR, plaque assay and indirect immunofluorescence. A total of nineteen methods were shown to reduce NiV infectious particles in cells, supernatants and organs to undetectable levels. Therefore, we provide a list of methods for the safe and efficient inactivation of NiV.
Infectivity and Morphology of Bovine Coronavirus Inactivated In Vitro by Cationic Photosensitizers
Authors: Zhukhovitsky Vladimir , Shevlyagina Natalia , Zubasheva Margarita , Russu Leonid , Gushchin Vladimir , Meerovich Gennady , Strakhovskaya Marina
Published: May 2022

Bovine coronaviruses (BCoVs), which cause gastrointestinal and respiratory diseases in cattle, and are genetically related to the human coronavirus HCoV-OC43, which is responsible for up to 10% of common colds, attract increased attention. We applied the method of photodynamic inactivation with cationic photosensitizers (PSs) to reduce the titers of BCoV and studied the morphological structure of viral particles under various modes of photodynamic exposure. The samples of virus containing liquid with an initial virus titer of 5 Log10 TCID50/mL were incubated with methylene blue (MB) or octakis(cholinyl)zinc phthalocyanine (Zn-PcChol8+) at concentrations of 1–5 μM for 10 min in the dark at room temperature. After incubation, samples were irradiated with LED (emission with maximum at 663 nm for MB or at 686 nm for Zn-PcChol8+) with light doses of 1.5 or 4 J/cm2. Next, the irradiation titrated virus containing liquid was studied using negative staining transmission electron microscopy. MB and Zn-PcChol8+ at concentrations of 1–5 μM, in combination with red light from LED sources in the low doses of 1.5–4.0 J/cm2, led to a decrease in BCoV titers by at least four orders of magnitude from the initial titer 5 Log10 TCID50/mL. Morphological changes in photodamaged BCoVs with increasing PS concentrations were loss of spikes, change in shape, decreased size of virus particles, destruction of the envelope, and complete disintegration of viruses. BCoV has been found to be sensitive to MB, which is the well-known approved drug, even in the absence of light.
Spread of COVID-19 Infection in Long-Term Care Facilities of Trieste (Italy) during the Pre-Vaccination Era, Integrating Findings of 41 Forensic Autopsies with Geriatric Comorbidity Index as a Valid Option for the Assessment of Strength of Causation
Authors: Zanon Martina , Peruch Michela , Concato Monica , Moreschi Carlo , Pizzolitto Stefano , Radaelli Davide , D’Errico Stefano
Published: May 2022

Background: in 2020, a new form of coronavirus spread around the world starting from China. The older people were the population most affected by the virus worldwide, in particular in Italy where more than 90% of deaths were people over 65 years. In these people, the definition of the cause of death is tricky due to the presence of numerous comorbidities. Objective: to determine whether COVID-19 was the cause of death in a series of older adults residents of nursing care homes. Methods: 41 autopsies were performed from May to June 2020. External examination, swabs, and macroscopic and microscopic examination were performed. Results: the case series consisted of nursing home guests; 15 men and 26 women, with a mean age of 87 years. The average number of comorbidities was 4. Based only on the autopsy results, the defined cause of death was acute respiratory failure due to diffuse alveolar damage (8%) or (31%) bronchopneumonia with one or more positive swabs for SARS-CoV-2. Acute cardiac failure with one or more positive swabs for SARS-CoV-2 was indicated as the cause of death in in symptomatic (37%) and asymptomatic (10%) patients. Few patients died for septic shock (three cases), malignant neoplastic diseases (two cases), and massive digestive bleeding (one case). Conclusions: Data from post-mortem investigation were integrated with previously generated Geriatric Index of Comorbidity (GIC), resulting in four different degrees of probabilities: high (12%), intermediate (10%), low (59%), and none (19%), which define the level of strength of causation and the role of COVID-19 disease in determining death.
Epitope Profiling of Diphtheria Toxoid Provides Enhanced Monitoring for Consistency Testing during Manufacturing Process Changes
Authors: Houy Camille , Ming Marin , Ettorre Luciano , Jin Robbie , Thangavadivel Nemika , Chen Tricia , Su Jin , Gajewska Beata
Published: May 2022

In the vaccine industry, multiple physicochemical, immunological, in vitro and in vivo analytical methods are applied throughout the manufacturing process to characterize and monitor the quality of vaccines. Presented here is the Single Epitope Antigenicity Test (SEAT), an innovative, quantitative epitope profiling method which provides an extended immunochemical analysis for diphtheria toxoid (DTxd) to be used for consistency testing during manufacturing process changes. The method uses BioLayer Interferometry (BLI) and a panel of monoclonal antibodies (mAbs) to independently assess nine individual antigenic sites of DTxd. The panel includes mAbs which are functional, bind distinct sites on DTxd and are able to distinguish intact DTxd from that which has been exposed to heat treatment. The SEAT method was qualified for precision, accuracy, and linearity, and was used to define a preliminary comparability range for DTxd made using the current manufacturing process. DTxd lots manufactured using alternate processes were assessed in the context of this range to determine the impact on DTxd antigenicity. Epitope profiling by SEAT provides quantitative information on the integrity of multiple important antigenic regions of DTxd, and therefore represents a valuable tool in a comprehensive analytical test package which can be used to support manufacturing process changes for vaccines.
Evaluation of Meningococcal Serogroup C Bactericidal Antibodies after Primary Vaccination: A Multicentre Study, Italy
Authors: Neri Arianna , Fabiani Massimo , Barbui Anna , Vocale Caterina , Miglietta Alessandro , Fazio Cecilia , Carannante Anna , Palmieri Annapina , Vacca Paola , Ambrosio Luigina , Stefanelli Paola
Published: May 2022

Here, we evaluated over time in different cohorts of children vaccinated against serogroup C Neisseria meningitidis, the presence of antibodies with neutralizing activity. A total of 348 sera samples of enrolled children by year since vaccination (<1 year- up to 5 years), starting from February 2016 to December 2017, were collected in three collaborating centers. Meningococcal serogroup C (MenC) antibody titers were measured with a serum bactericidal antibody (SBA) assay using rabbit complement (rSBA) following standard operating procedures. The cut-off of rSBA titer ≥ 8 is considered the correlate of protection. We observed a significantly declining of bactericidal rSBA titers by 23% every year, for every 1-year from vaccination (Adjusted PR = 0.77, 95% CI: 0.71–0.84). The proportions of children with bactericidal antibodies, immunized with the meningococcal serogroup C conjugate (MCC) vaccine, declined from 67.7% (95% CI: 48.6–83.3%) one year after vaccination, to 36.7% (95% CI: 19.9–56.1%) five years after vaccination (chi-square for linear trend, p < 0.001). Children vaccinated with the tetravalent meningococcal serogroup ACWY vaccine resulted in a high proportion of bactericidal rSBA MenC titer ≥ 1:8 (90.6%, 95% CI: 79.3–96.9%) after a mean time of seven months. Overall, the results provide some evidences on the evaluation of meningococcal serogroup C bactericidal antibodies after primary vaccination.
Real-World Effectiveness of the mRNA COVID-19 Vaccines in Japan: A Case–Control Study
Authors: Hara Megumi , Furue Takeki , Fukuoka Mami , Iwanaga Kentaro , Matsuishi Eijo , Miike Toru , Sakamoto Yuichiro , Mukai Naoko , Kinugasa Yuki , Shigyo Mutsumi , Sonoda Noriko , Tanaka Masato , Arase Yasuko , Tanaka Yosuke , Nakashima Hitoshi , Irie Shin , Hirota Yoshio
Published: May 2022

The real-world effectiveness of the coronavirus disease 2019 (COVID-19) vaccines in Japan remains unclear. This case–control study evaluated the vaccine effectiveness (VE) of two doses of mRNA vaccine, BNT162b2 or mRNA-1273, against the delta (B.1.617.2) variant in the Japanese general population in the period June–September 2021. Individuals in close contact with COVID-19 patients were tested using polymerase chain reaction (PCR). A self-administered questionnaire evaluated vaccination status, demographic data, underlying medical conditions, lifestyle, personal protective health behaviors, and living environment. Two vaccine doses were reported by 11.6% of cases (n = 389) and 35.2% of controls (n = 179). Compared with controls, cases were younger and had a lower proportion who always performed handwashing for ≥20 s, a higher proportion of alcohol consumers, and a lower proportion of individuals living in single-family homes or with commuting family members. After adjusting for these confounding factors and day of PCR testing by multivariate logistic regression analysis, the VE in the period June–July (delta variant proportion 45%) was 92% and 79% in the period August–September (delta variant proportion 89%). The adjusted VE for homestay, hotel-based isolation and quarantine, and hospitalization was 78%, 77%, and 97%, respectively. Despite declining slightly, VE against hospitalization remained robust for ~3 months after the second dose. Vaccination policymaking will require longer-term monitoring of VE against new variants.
Factors Influencing COVID-19 Vaccine Uptake among Nepali People in the UK: A Qualitative Study
Authors: Simkhada Padam , Tamang Pasang , Timilsina Laxmi , Simkhada Bibha , Bissell Paul , van Teijlingen Edwin , Sah Sunil Kumar , Wasti Sharada Prasad
Published: May 2022

Vaccination saves lives and can be an effective strategy for preventing the spread of the COVID-19, but negative attitudes towards vaccines lead to vaccine hesitancy. This study aimed to explore the factors influencing the uptake of the COVID-19 vaccine in the Nepali community in the United Kingdom (UK). This qualitative study included in-depth interviews with 20 people from Nepal living in the UK. Interviews were conducted by a native-Nepali speaker and all interviews were audio-recorded, transcribed, and translated into English before being analysed thematically. Our study found that attitudes towards COVID-19 are generally positive. Nine overlapping themes around barriers to COVID-19 vaccination were identified: (a) rumours and mis/disinformation; (b) prefer home remedies and yoga; (c) religion restriction; (d) concern towards vaccine eligibility; (e) difficulty with online vaccine booking system; (f) doubts of vaccine effectiveness after changing the second dose timeline; (g) lack of confidence in the vaccine; (h) past bad experience with the influenza vaccine; and (i) worried about side-effects. Understanding barriers to the uptake of the COVID-19 vaccine can help in the design of better targeted interventions. Public health messages including favourable policy should be tailored to address those barriers and make this vaccination programme more viable and acceptable to the ethnic minority communities in the UK.