bluuebunny/crossref_metadata_2025 huggingface dataset:
Comparison of Indoor Air Quality in Summer and Winter According to Four-Way Cassette Fan Coil Unit Operation in a Four-Bed Ward
Authors: Lee Jungsuk , An Ik-Hyun , Park Su-Hoon , Lee Kyung-Rae , Kim Young-Won , Yook Se-Jin
Published: August 2022

This study targeted a four-bed ward with a ventilation system and a four-way cassette fan coil unit (4-way FCU) installed on the ceiling. The indoor air quality under summer and winter conditions was comparatively analyzed. The age of air was calculated by conducting tests and simulations under diverse conditions, assuming that the ventilation system and 4-way FCU were continuously operating. The use of an air cleaner and ward curtain was investigated for its impact on the air quality in the breathing zone of a patient lying on the bed, and effects of the airflow and discharge angle of the 4-way FCU were considered. Because the 4-way FCU was installed in the central part of the ceiling, where indoor air is sucked in and subsequently discharged in four directions, the age of air at each bed was found to vary depending on the airflow and discharge angle of the 4-way FCU. When the airflow and discharge angle of the 4-way FCU was fixed, the age of air at each bed appeared to be lower during winter heating than in summer cooling mode. The age of air was significantly lowered at each bed, depending on the use of the curtain and the air cleaner along with the ventilation system and 4-way FCU, and appropriate seasonal operating conditions were identified to maintain a lower age of air at each bed.
A Critical Review of Recent Literature on Metal Contents in E-Cigarette Aerosol
Authors: Soulet Sebastien , Sussman Roberto A.
Published: August 2022

The inhalation of metallic compounds in e-cigarette (EC) aerosol emissions presents legitimate concerns of potential harms for users. We provide a critical review of laboratory studies published after 2017 on metal contents in EC aerosol, focusing on the consistency between their experimental design, real life device usage and appropriate evaluation of exposure risks. All experiments reporting levels above toxicological markers for some metals (e.g., nickel, lead, copper, manganese) exhibited the following experimental flaws: (i) high powered sub-ohm tank devices tested by means of puffing protocols whose airflows and puff volumes are conceived and appropriate for low powered devices; this testing necessarily involves overheating conditions that favor the production of toxicants and generate aerosols that are likely repellent to human users; (ii) miscalculation of exposure levels from experimental outcomes; (iii) pods and tank devices acquired months and years before the experiments, so that corrosion effects cannot be ruled out; (iv) failure to disclose important information on the characteristics of pods and tank devices, on the experimental methodology and on the resulting outcomes, thus hindering the interpretation of results and the possibility of replication. In general, low powered devices tested without these shortcomings produced metal exposure levels well below strict reference toxicological markers. We believe this review provides useful guidelines for a more objective risk assessment of EC aerosol emissions and signals the necessity to upgrade current laboratory testing standards.
Safety Profile and Lack of Immunogenicity of IncobotulinumtoxinA in Pediatric Spasticity and Sialorrhea: A Pooled Analysis
Authors: Berweck Steffen , Banach Marta , Gaebler-Spira Deborah , Chambers Henry G. , Schroeder A. S. , Geister Thorin L. , Althaus Michael , Hanschmann Angelika , Vacchelli Matteo , Bonfert Michaela V. , Heinen Florian , Dabrowski Edward
Published: August 2022

IncobotulinumtoxinA, a pure botulinumtoxinA formulation, is free of accessory proteins. This analysis provides pooled safety data from phase 3 trials of children/adolescents (2–17 years), investigating incobotulinumtoxinA for the treatment of spasticity associated with cerebral palsy (at doses ≤20 U/kg (max. 500 U) per injection cycle (IC) for ≤6 ICs; three trials) or sialorrhea associated with neurologic disorders (at total doses of 20–75 U per IC for ≤4 ICs; one trial) for ≤96 weeks. Safety endpoints included the incidences of different types of treatment-emergent adverse events (TEAEs) and immunogenicity. IncobotulinumtoxinA dose groups were combined. Of 1159 patients (mean age 7.3 years, 60.4% males) treated with incobotulinumtoxinA, 3.9% experienced treatment-related TEAEs, with the most common being injection site reactions (1.3%) (both indications), muscular weakness (0.7%) (spasticity), and dysphagia (0.2%) (sialorrhea). Two patients (0.2%) experienced a treatment-related treatment-emergent serious adverse event, and 0.3% discontinued the study due to treatment-related TEAEs. No botulinumtoxinA-naïve patients developed neutralizing antibodies (NAbs) after incobotulinumtoxinA. All children/adolescents with known pre-treatment status and testing positive for Nabs at final visit (n = 7) were previously treated with a botulinumtoxinA other than incobotulinumtoxinA. IncobotulinumtoxinA was shown to be safe, with very few treatment-related TEAEs in a large, diverse cohort of children/adolescents with chronic conditions requiring long-term treatment and was without new NAb formation in treatment-naïve patients.
The Strong Anti-Tumor Effect of Smp24 in Lung Adenocarcinoma A549 Cells Depends on Its Induction of Mitochondrial Dysfunctions and ROS Accumulation
Authors: Guo Ruiyin , Chen Xuewen , Nguyen Tienthanh , Chai Jinwei , Gao Yahua , Wu Jiena , Li Jinqiao , Abdel-Rahman Mohamed A. , Chen Xin , Xu Xueqing
Published: August 2022

Non-small cell lung cancer (NSCLC) is the leading cause of death in lung cancer due to its aggressiveness and rapid migration. The potent antitumor effect of Smp24, an antimicrobial peptide derived from Egyptian scorpion Scorpio maurus palmatus via damaging the membrane and cytoskeleton have been reported earlier. However, its effects on mitochondrial functions and ROS accumulation in human lung cancer cells remain unknown. In the current study, we discovered that Smp24 can interact with the cell membrane and be internalized into A549 cells via endocytosis, followed by targeting mitochondria and affect mitochondrial function, which significantly causes ROS overproduction, altering mitochondrial membrane potential and the expression of cell cycle distribution-related proteins, mitochondrial apoptotic pathway, MAPK, as well as PI3K/Akt/mTOR/FAK signaling pathways. In summary, the antitumor effect of Smp24 against A549 cells is related to the induction of apoptosis, autophagy plus cell cycle arrest via mitochondrial dysfunction, and ROS accumulation. Accordingly, our findings shed light on the anticancer mechanism of Smp24, which may contribute to its further development as a potential agent in the treatment of lung cancer cells.
Gut Non-Bacterial Microbiota: Emerging Link to Irritable Bowel Syndrome
Authors: Liu Ao , Gao Wenkang , Zhu Yixin , Hou Xiaohua , Chu Huikuan
Published: August 2022

As a common functional gastrointestinal disorder, irritable bowel syndrome (IBS) significantly affects personal health and imposes a substantial economic burden on society, but the current understanding of its occurrence and treatment is still inadequate. Emerging evidence suggests that IBS is associated with gut microbial dysbiosis, but most studies focus on the bacteria and neglect other communities of the microbiota, including fungi, viruses, archaea, and other parasitic microorganisms. This review summarizes the latest findings that link the nonbacterial microbiota with IBS. IBS patients show less fungal and viral diversity but some alterations in mycobiome, virome, and archaeome, such as an increased abundance of Candida albicans. Moreover, fungi and methanogens can aid in diagnosis. Fungi are related to distinct IBS symptoms and induce immune responses, intestinal barrier disruption, and visceral hypersensitivity via specific receptors, cells, and metabolites. Novel therapeutic methods for IBS include fungicides, inhibitors targeting fungal pathogenic pathways, probiotic fungi, prebiotics, and fecal microbiota transplantation. Additionally, viruses, methanogens, and parasitic microorganisms are also involved in the pathophysiology and treatment. Therefore, the gut nonbacterial microbiota is involved in the pathogenesis of IBS, which provides a novel perspective on the noninvasive diagnosis and precise treatment of this disease.
In Silico Evaluation of Sesquiterpenes and Benzoxazinoids Phytotoxins against Mpro, RNA Replicase and Spike Protein of SARS-CoV-2 by Molecular Dynamics. Inspired by Nature
Authors: Mejías Francisco J. R. , Durán Alexandra G. , Chinchilla Nuria , Varela Rosa M. , Álvarez José A. , Molinillo José M. G. , García-Cozar Francisco , Macías Francisco A.
Published: August 2022

In the work described here, a number of sesquiterpenes and benzoxazinoids from natural sources, along with their easily accessible derivatives, were evaluated against the main protease, RNA replicase and spike glycoprotein of SARS-CoV-2 by molecular docking. These natural products and their derivatives have previously shown remarkable antiviral activities. The most relevant compounds were the 4-fluoro derivatives of santamarine, reynosin and 2-amino-3H-phenoxazin-3-one in terms of the docking score. Those compounds fulfill the Lipinski’s rule, so they were selected for the analysis by molecular dynamics, and the kinetic stabilities of the complexes were assessed. The addition of the 4-fluorobenzoate fragment to the natural products enhances their potential against all of the proteins tested, and the complex stability after 50 ns validates the inhibition calculated. The derivatives prepared from reynosin and 2-amino-3H-phenoxazin-3-one are able to generate more hydrogen bonds with the Mpro, thus enhancing the stability of the protein–ligand and generating a long-term complex for inhibition. The 4-fluoro derivate of santamarine and reynosin shows to be really active against the spike protein, with the RMSD site fluctuation lower than 1.5 Å. Stabilization is mainly achieved by the hydrogen-bond interactions, and the stabilization is improved by the 4-fluorobenzoate fragment being added. Those compounds tested in silico reach as candidates from natural sources to fight this virus, and the results concluded that the addition of the 4-fluorobenzoate fragment to the natural products enhances their inhibition potential against the main protease, RNA replicase and spike protein of SARS-CoV-2.
µ-Conotoxins Targeting the Human Voltage-Gated Sodium Channel Subtype NaV1.7
Authors: McMahon Kirsten L. , Tran Hue N. T. , Deuis Jennifer R. , Craik David J. , Vetter Irina , Schroeder Christina I.
Published: August 2022

µ-Conotoxins are small, potent, peptide voltage-gated sodium (NaV) channel inhibitors characterised by a conserved cysteine framework. Despite promising in vivo studies indicating analgesic potential of these compounds, selectivity towards the therapeutically relevant subtype NaV1.7 has so far been limited. We recently identified a novel µ-conotoxin, SxIIIC, which potently inhibits human NaV1.7 (hNaV1.7). SxIIIC has high sequence homology with other µ-conotoxins, including SmIIIA and KIIIA, yet shows different NaV channel selectivity for mammalian subtypes. Here, we evaluated and compared the inhibitory potency of µ-conotoxins SxIIIC, SmIIIA and KIIIA at hNaV channels by whole-cell patch-clamp electrophysiology and discovered that these three closely related µ-conotoxins display unique selectivity profiles with significant variations in inhibitory potency at hNaV1.7. Analysis of other µ-conotoxins at hNaV1.7 shows that only a limited number are capable of inhibition at this subtype and that differences between the number of residues in loop 3 appear to influence the ability of µ-conotoxins to inhibit hNaV1.7. Through mutagenesis studies, we confirmed that charged residues in this region also affect the selectivity for hNaV1.4. Comparison of µ-conotoxin NMR solution structures identified differences that may contribute to the variance in hNaV1.7 inhibition and validated the role of the loop 1 extension in SxIIIC for improving potency at hNaV1.7, when compared to KIIIA. This work could assist in designing µ-conotoxin derivatives specific for hNaV1.7.
Prevalence and Antifungal Susceptibility of Clinically Relevant Candida Species, Identification of Candida auris and Kodamaea ohmeri in Bangladesh
Authors: Sathi Fardousi Akter , Paul Shyamal Kumar , Ahmed Salma , Alam Mohammad Monirul , Nasreen Syeda Anjuman , Haque Nazia , Islam Arup , Nila Sultana Shabnam , Afrin Sultana Zahura , Aung Meiji Soe , Kobayashi Nobumichi
Published: August 2022

Candida species are major fungal pathogens in humans. The aim of this study was to determine the prevalence of individual Candida species and their susceptibility to antifungal drugs among clinical isolates in a tertiary care hospital in Bangladesh. During a 10-month period in 2021, high vaginal swabs (HVSs), blood, and aural swabs were collected from 360 patients. From these specimens, Candida spp. was isolated from cultures on Sabouraud dextrose agar media, and phenotypic and genetic analyses were performed. A total of 109 isolates were recovered, and C. albicans accounted for 37%, being derived mostly from HVSs. Among non-albicans Candida (NAC), C. parapsilosis was the most frequent, followed by C. ciferrii, C. tropicalis, and C. glabrata. Three isolates from blood and two isolates from aural discharge were genetically identified as C. auris and Kodamaea ohmeri, respectively. NAC isolates were more resistant to fluconazole (overall rate, 29%) than C. albicans (10%). Candida isolates from blood showed 95% susceptibility to voriconazole and less susceptibility to fluconazole (67%). Two or three amino acid substitutions were detected in the ERG11 of two fluconazole-resistant C. albicans isolates. The present study is the first to reveal the prevalence of Candida species and their antifungal susceptibility in Bangladesh.
Assessing Public Awareness of the Malaria Vaccine in Sub-Saharan Africa
Authors: Aremu Taiwo Opeyemi , Singhal Chinar , Ajibola Oluwafemi Augustine , Agyin-Frimpong Emmanuel , Appiah-Num Safo Akua Asantewaa , Ihekoronye Maduabuchi Romanus , Nabirye Stella Esther , Okoro Olihe Nnenna
Published: August 2022

Background: Malaria infection remains one of the leading causes of death in sub-Saharan Africa. Over the years, several measures have been implemented for the prevention of malaria, including vector control with insecticide-treated nets, indoor residual spraying, and seasonal or traveling prophylactics. In 2021, the World Health Organization (WHO) approved the use of the malaria vaccine in children. We conducted a cross-sectional survey study in three sub-Saharan African countries—Uganda, Ghana, and Nigeria—to assess public awareness of the malaria vaccine among the residents of these countries. Method: A cross-sectional, web-based survey was conducted between time January 2022 and April 2022 using Qualtrics® software (Version number: April 2022; Qualtrics, Provo, UT, USA). A total of 3896 responses were analyzed using SAS OnDemand for Academics software. Linear regression model was used to assess the relationship between the demographic characteristics and awareness of the malaria vaccine, using a level of significance (alpha) of 0.05. Result: Overall, there was significant association between the level of education and public awareness of the malaria vaccine in each of the countries studied. Gender and place of residence were associated with awareness in Nigeria and Uganda, while younger respondents were more likely to be aware of the malaria vaccine in Ghana. Conclusion: Given the negative impact of lack of awareness and knowledge, misinformation and conspiracy theories on immunization programs, public health campaigns preceding the population-wide roll-out of the novel malaria vaccine should target the less-educated, and those residing in more rural areas, while assuring equitable access to the malaria vaccine across sub-Saharan Africa.
Three-Dimensional Models of Soil-Transmitted Helminth Eggs from Light Microscopy Images
Authors: Dias Yan Emygdio , de Freitas Elisângela Oliveira , de Oliveira Dayane Alvarinho , Girard-Dias Wendell , Crivano Machado Lúcio Paulo do Amaral , Lopes-Torres Eduardo José
Published: August 2022

The World Health Organization indicates that more than 1.5 billion people are infected with geohelminths. Soil-transmitted helminths prevail mostly in tropical and subtropical regions, in areas with inadequate hygiene and sanitation conditions, and basic health education problems. Nematode eggs are structures of resistance and infection by fecal–oral transmission. When STH eggs are ingested, they can infect the potential host, causing abdominal pain, diarrhea, anemia, malnutrition, and physical-cognitive impacts in children. Taking advantage of the increasing employment of three-dimensional models of these structured based on light microscopy images to improve the research area and education could be an alternative to improve health education and spread scientific information on transmission and prevention. The objective of this work was to produce 3D printed models from bi-dimensional images of eggs based on their real morphological and morphometric characteristics. The virtual models were reconstructed from the acquisition and selection of images obtained using light microscopy. After selecting referential images, we constructed the models based on the vectorization of the egg structures. After vectorization, 3D modeling was performed and printed in PLA. 3D models have a high potential to contribute to the advanced morphological studies and teaching of parasitological sciences, enriching the teaching-learning process applicable in presential or remote teaching of basic education, undergraduate, and post-graduation classes.
Prevalence of Escherichia coli ST1193 Causing Intracranial Infection in Changsha, China
Authors: Zhong Yi-Ming , Zhang Xiao-He , Ma Zheng , Liu Wen-En
Published: August 2022

ST1193 is an emerging new virulent and resistant clone among Escherichia coli with a tendency to spread rapidly across the globe. However, the prevalence of intracranial infection-causing E. coli ST1193 is rarely reported. This study aimed at determining the prevalence of E. coli ST1193 isolates, causing intracranial infections in Changsha, central China. A total of 28 E. coli isolates were collected from the cerebrospinal fluid of patients with intracranial infection over a four-year period. All isolates were differentiated using multilocus sequence typing (MLST), and phylogenetic grouping, and tested for antibiotic resistance. MLST analysis showed 11 sequence types (ST) among the 28 E. coli isolates. The most prevalent ST was B2-ST1193 (28.6%, 8/28), followed by B2-ST131 (21.4%, 6/28) and F-ST648 (10.7%, 3/28). Of the eight ST1193 isolates, three carried CTX-M-55, and one carried CTX-M-27. All eight ST1193 isolates were resistant to Ciprofloxacin, showing gyrA1AB/parC4A mutations. Two ST1193 isolates carried the aac(6′)-Ib-cr gene. All ST1193 isolates were recovered from infants with meningitis, with a fatal outcome for one three-month-old infant. ST1193 has emerged as the predominant type of E. coli strain causing intracranial infections in Changsha, China. This study highlights the importance of implementing appropriate surveillance measures to prevent the spread of this emerging public health threat.
Search for Gravitational-Neutrino Correlations on Ground-Based Detectors
Authors: Andrusenko Svetlana , Gavriluk Yurii , Gusev Andrei , Krichevskiy Daniil , Oreshkin Sergei , Popov Sergei , Rudenko Valentin
Published: August 2022

The problem of joint data processing from ground-based gravitational and neutrino detectors is considered in order to increase the detection efficiency of collapsing objects in the Galaxy. The development of the “neutrino-gravitational correlation” algorithm is carried out within the framework of the theory of optimal filtration as applied to the well-known OGRAN and BUST facilities located at the BNO INR RAS. The experience of analyzing neutrino and gravitational data obtained during the outburst of supernova SN1987A is used. Sequential steps of the algorithm are presented; formulas for estimating the statistical efficiency of a two-channel recorder are obtained.
Cosmology from Strong Interactions
Authors: Addazi Andrea , Lundberg Torbjörn , Marcianò Antonino , Pasechnik Roman , Šumbera Michal
Published: August 2022

The wealth of theoretical and phenomenological information about Quantum Chromodynamics at short and long distances collected so far in major collider measurements has profound implications in cosmology. We provide a brief discussion on the major implications of the strongly coupled dynamics of quarks and gluons as well as on effects due to their collective motion on the physics of the early universe and in astrophysics.
A Study on the Promotional Effect and Mechanism of National e-Commerce Demonstration City Construction on Green Innovation Capacity of Cities
Authors: Li Jie , Yuan Shengjun , Wu Jun
Published: August 2022

Green innovation precisely reflects China’s new development concept and is the general trend for urban development in the future. In addition, e-commerce might become an important breakthrough platform to promote urban green innovation. In order to explore the impact of the construction of national e-commerce demonstration cities on urban green innovation, this paper aimed to theoretically analyze the potential relationship between the two and the transmission mechanism. At the same time, we have examined the panel data from 297 prefecture level cities in China from 2005 to 2018 to explore the implementation effect of the pilot policies by using difference-in-differences and carried out a series of robustness tests. The results showed that: (i) The pilot policy of e-commerce demonstration cities exhibited a significant promotion effect on the green innovation capacity of the different cities, and in general, the promotion effect of the pilot policy is dynamically sustainable. (ii) Analysis of the further influence mechanism showed that the pilot policy could effectively promote the development of urban green innovation capacity by enhancing the level of urban informatization, thereby attracting the concentration of scientific and technological talents. This in turn can facilitate urban innovation and the entrepreneurship environment, among which the boosting effect of optimizing the urban innovation and entrepreneurship environment was the greatest. (iii) In terms of heterogeneity, the pilot policies showed significant positive effects on all the regions to which the cities belong, while the boosting effects were more significant for cities with large populations, non-central cities, and the general science and education cities. The findings of this study not only enrich the research results in the field of urban green innovation, but also have clear policy implications, which can provide useful guidance and reference value for the work of relevant departments.
Seed Transmission of Tomato Spotted Wilt Orthotospovirus in Peppers
Authors: Wang Hongwei , Wu Xiujuan , Huang Xiande , Wei Shujun , Lu Zhijun , Ye Jian
Published: August 2022

Tomato spotted wilt orthotospovirus (TSWV) severely damaged agricultural production in many places around the world. It is generally believed that TSWV transmits among plants via their insect vector. In this study, we provide evidence on the seed-borne transmission of TSWV in pepper (Capsicum annuum L.) plants. RT-PCR, RT-qPCR, and transmission electron microscopy data demonstrate the seed transmission ability of TSWV in peppers. Endosperm, but not the embryo, is the abundant virus-containing seed organ. TSWV can also be detected in the second generation of newly germinated seedlings from virus-containing seed germination experiments. Our data are useful for researchers, certification agencies, the seed industry, and policy makers when considering the importance of TSWV in vegetable production all over the world.
Cellular Lipids—Hijacked Victims of Viruses
Authors: Omasta Bozena , Tomaskova Jana
Published: August 2022

Over the millions of years-long co-evolution with their hosts, viruses have evolved plenty of mechanisms through which they are able to escape cellular anti-viral defenses and utilize cellular pathways and organelles for replication and production of infectious virions. In recent years, it has become clear that lipids play an important role during viral replication. Viruses use cellular lipids in a variety of ways throughout their life cycle. They not only physically interact with cellular membranes but also alter cellular lipid metabolic pathways and lipid composition to create an optimal replication environment. This review focuses on examples of how different viruses exploit cellular lipids in different cellular compartments during their life cycles.
Virus Diversity, Abundance, and Evolution in Three Different Bat Colonies in Switzerland
Authors: Wiederkehr Myriam Anja , Qi Weihong , Schoenbaechler Katja , Fraefel Cornel , Kubacki Jakub
Published: August 2022

Bats are increasingly recognized as reservoirs for many different viruses that threaten public health, such as Hendravirus, Ebolavirus, Nipahvirus, and SARS- and MERS-coronavirus. To assess spillover risk, viromes of bats from different parts of the world have been investigated in the past. As opposed to most of these prior studies, which determined the bat virome at a single time point, the current work was performed to monitor changes over time. Specifically, fecal samples of three endemic Swiss bat colonies consisting of three different bat species were collected over three years and analyzed using next-generation sequencing. Furthermore, single nucleotide variants of selected DNA and RNA viruses were analyzed to investigate virus genome evolution. In total, sequences of 22 different virus families were found, of which 13 are known to infect vertebrates. Most interestingly, in a Vespertilio murinus colony, sequences from a MERS-related beta-coronavirus were consistently detected over three consecutive years, which allowed us to investigate viral genome evolution in a natural reservoir host.
Vertical Transmission of Sindbis Virus in Culex Mosquitoes
Authors: Dahl Emma , Öborn Linnea , Sjöberg Viktoria , Lundkvist Åke , Hesson Jenny C.
Published: August 2022

Vertical transmission (VT) is a phenomenon of vector-borne diseases where a pathogen is transferred from an infected arthropod mother to her offspring. For mosquito-borne flavi- and alphaviruses, VT is commonly viewed as rare; however, both field and experimental studies report on vertical transmission efficiency to a notably varying degree. It is likely that this reflects the different experimental methods used to test vertical transmission efficiency as well as differences between virus–vector combinations. There are very few investigations of the VT of an alphavirus in a Culex vector. Sindbis virus (SINV) is an arthritogenic alphavirus that utilizes Culex species as main vectors both in the summer transmission season and for its persistence over the winter period in northern latitudes. In this study, we investigated the vertical transmission of the SINV in Culex vectors, both in the field and in experimental settings. The detection of SINV RNA in field-collected egg rafts and emerging adults shows that vertical transmission takes place in the field. Experimentally infected females gave rise to adult offspring containing SINV RNA at emergence; however, three to four weeks after emergence none of the offspring contained SINV RNA. This study shows that vertical transmission may be connected to SINV’s ability to persist throughout northern winters and also highlights many aspects of viral replication that need further study.
Deletion of the CD2v Gene from the Genome of ASFV-Kenya-IX-1033 Partially Reduces Virulence and Induces Protection in Pigs
Authors: Hemmink Johanneke D. , Khazalwa Emmanuel M. , Abkallo Hussein M. , Oduor Bernard , Khayumbi Jeremiah , Svitek Nicholas , Henson Sonal P. , Blome Sandra , Keil Günther , Bishop Richard P. , Steinaa Lucilla
Published: August 2022

Infection of pigs with the African swine fever virus (ASFV) leads to a devastating hemorrhagic disease with a high mortality of up to 100%. In this study, a CD2v gene deletion was introduced to a genotype IX virus from East Africa, ASFV-Kenya-IX-1033 (ASFV-Kenya-IX-1033-∆CD2v), to investigate whether this deletion led to reduced virulence in domestic pigs and to see if inoculation with this LA-ASFV could induce protective immunity against parental virus challenge. All pigs inoculated with ASFV-Kenya-IX-1033-ΔCD2v survived inoculation but presented with fever, reduced appetite and lethargy. ASFV genomic copies were detected in only one animal at one time point. Seven out of eight animals survived subsequent challenge with the pathogenic parental strain (87.5%) but had mild to moderate clinical symptoms and had a gross pathology compatible with chronic ASFV infection. All mock-immunised animals developed acute ASF upon challenge with ASFV-Kenya-IX-1033 and were euthanised upon meeting the humane endpoint criteria. ASFV genome copy numbers after challenge were similar in the two groups. ASFV-Kenya-IX-1033-∆CD2v is therefore a useful tool to investigate the development of immunity to ASFV genotype IX, but safety concerns preclude its use as a candidate vaccine without further attenuation.
Structural Dynamics and Activity of B19V VP1u during the pHs of Cell Entry and Endosomal Trafficking
Authors: Lakshmanan Renuk V. , Hull Joshua A. , Berry Luke , Burg Matthew , Bothner Brian , McKenna Robert , Agbandje-McKenna Mavis
Published: August 2022

Parvovirus B19 (B19V) is a human pathogen that is the causative agent of fifth disease in children. It is also known to cause hydrops in fetuses, anemia in AIDS patients, and transient aplastic crisis in patients with sickle cell disease. The unique N-terminus of Viral Protein 1 (VP1u) of parvoviruses, including B19V, exhibits phospholipase A2 (PLA2) activity, which is required for endosomal escape. Presented is the structural dynamics of B19V VP1u under conditions that mimic the pHs of cell entry and endosomal trafficking to the nucleus. Using circular dichroism spectroscopy, the receptor-binding domain of B19V VP1u is shown to exhibit an α-helical fold, whereas the PLA2 domain exhibits a probable molten globule state, both of which are pH invariant. Differential scanning calorimetry performed at endosomal pHs shows that the melting temperature (Tm) of VP1u PLA2 domain is tuned to body temperature (37 °C) at pH 7.4. In addition, PLA2 assays performed at temperatures ranging from 25–45 °C show both a temperature and pH-dependent change in activity. We hypothesize that VP1u PLA2 domain differences in Tm at differing pHs have enabled the virus to “switch on/off” the phospholipase activity during capsid trafficking. Furthermore, we propose the environment of the early endosome as the optimal condition for endosomal escape leading to B19V infection.
Full-Length Genomic RNA of Foot-and-Mouth Disease Virus Is Infectious for Cattle by Injection
Authors: Keck Hanna , Litz Benedikt , Hoffmann Bernd , Sehl-Ewert Julia , Beer Martin , Eschbaumer Michael
Published: August 2022

Safe sample transport is of great importance for infectious diseases diagnostics. Various treatments and buffers are used to inactivate pathogens in diagnostic samples. At the same time, adequate sample preservation, particularly of nucleic acids, is essential to allow an accurate laboratory diagnosis. For viruses with single-stranded RNA genomes of positive polarity, such as foot-and-mouth disease virus (FMDV), however, naked full-length viral RNA can itself be infectious. In order to assess the risk of infection from inactivated FMDV samples, two animal experiments were performed. In the first trial, six cattle were injected with FMDV RNA (isolate A22/IRQ/24/64) into the tongue epithelium. All animals developed clinical disease within two days and FMDV was reisolated from serum and saliva samples. In the second trial, another group of six cattle was exposed to FMDV RNA by instilling it on the tongue and spraying it into the nose. The animals were observed for 10 days after exposure. All animals remained clinically unremarkable and virus isolation as well as FMDV genome detection in serum and saliva were negative. No transfection reagent was used for any of the animal inoculations. In conclusion, cattle can be infected by injection with naked FMDV RNA, but not by non-invasive exposure to the RNA. Inactivated FMDV samples that contain full-length viral RNA carry only a negligible risk of infecting animals.
Designing a Multi-Epitope Vaccine against Toxoplasma gondii: An Immunoinformatics Approach
Authors: Hammed-Akanmu Mutiat , Mim Maria , Osman Abdinasir Yusuf , Sheikh Abdulrahman M. , Behmard Esmaeil , Rabaan Ali A. , Suppain Rapeah , Hajissa Khalid
Published: August 2022

Infection with the intracellular apicomplexan parasite Toxoplasma gondii causes serious clinical outcomes in both human and veterinary settings worldwide. Although approximately one-third of the world’s population is infected with T. gondii, an effective human vaccine for this disease remains unavailable. We aimed to design a potential T. gondii vaccine candidate that consisted of the B- and T-lymphocyte epitopes of three parasite immunogenic antigens. Firstly, the immunodominant epitopes expressed within the ROP2, MIC3, and GRA7 proteins of T. gondii were identified. Subsequently, six B-cell epitopes, five CTL epitopes, and five HTL epitopes were combined to generate a multi-epitope vaccine, and the 50S ribosomal protein L7/L12 was added as an adjuvant to boost the vaccine’s immunogenicity. All these epitopes were found to be antigenic, nonallergenic, nontoxic, and nonhuman homologs. The designed vaccine construct has a molecular weight of 51 kDa, an antigenicity score of 0.6182, and a solubility of 0.903461. Likewise, the candidate vaccine was immunogenic, nonallergenic, and stable. Molecular docking analysis revealed stable interactions between the vaccine construct and the TLR-4 immune receptor. Meanwhile, the stability of the developed vaccine was validated using molecular dynamics simulation. In silico, the vaccine construct was able to trigger primary immune responses. However, further laboratory-based assessments are needed to confirm its efficacy and safety.
An O-Specific Polysaccharide/Tetanus Toxoid Conjugate Vaccine Induces Protection in Guinea Pigs against Virulent Challenge with Coxiella burnetii
Authors: Graves Stephen R. , Islam Aminul , Webb Lawrence D. , Marsh Ian , Plain Karren , Westman Mark , Conlan Xavier A. , Carbis Rodney , Toman Rudolf , Stenos John
Published: August 2022

Q fever is caused by the bacterium Coxiella burnetii and is spread to humans from infected animals especially goats, sheep and cattle, predominantly when giving birth. There is an effective human vaccine (Q-VAX) against Q fever, and although Q fever is a worldwide problem, the vaccine is only used in Australia due to difficulties associated with its use and the risk of adverse reactions. The desire to protect humans, particularly farmers and abattoir workers, from Q fever prompted the development of a new safe and effective human vaccine without all the difficulties associated with the current vaccine. Candidate vaccines were prepared using purified O-specific polysaccharide (OSP) extracted from the lipopolysaccharide of virulent (phase 1) C. burnetii, strain Nine Mile, which was then conjugated to a tetanus toxoid (TT) carrier protein. Two vaccines were prepared using OSP from C. burnetii grown in embryonated eggs (vaccine A) and axenic media (vaccine B). Vaccines with or without alum adjuvant were used to vaccinate guinea pigs, which were later challenged by intranasal inoculation with virulent C. burnetii. Both vaccines protected guinea pigs from fever and loss of weight post challenge. Post-mortem samples of the spleen, liver and kidney of vaccinated guinea pigs contained substantially less C. burnetii DNA as measured by PCR than those of the unvaccinated control animals. This study demonstrated that a C. burnetii OSP-TT conjugate vaccine is capable of inducing protection against virulent C. burnetii in guinea pigs. Additionally, OSP derived from C. burnetii grown in axenic media compared to OSP from embryonated eggs is equivalent in terms of providing a protective immune response.
Factors Driving COVID-19 Vaccine Hesitancy in Cameroon and Their Implications for Africa: A Comparison of Two Cross-Sectional Studies Conducted 19 Months Apart in 2020 and 2022
Authors: Dinga Jerome Nyhalah , Njoh Andreas Ateke , Gamua Stanley Dobgima , Muki Synthia Eni , Titanji Vincent P. K.
Published: August 2022

Many efficacious COVID-19 vaccines have been approved for general use but their ability to control the disease is being undermined by slow uptake. Resources are needed to persuade people to obtain a COVID-19 vaccine. Here we compare this present study and a previous one to assess the impact of the Cameroon government’s policy and efforts to reduce COVID-19 vaccine hesitancy after one year of implementation. After obtaining ethical clearance and informed consent, 6732 participants completed a questionnaire about COVID-19 vaccine hesitancy and acceptance. It was observed that the government’s policies and efforts reduced COVID-19 vaccine hesitancy significantly, but this was not enough to ensure the herd immunity necessary to control the disease. The risk factors associated with vaccine hesitancy were the consumption of traditional herbal remedies; living in an urban setting; being female, jobless or a student; working in the education sector; being a politician/policy maker/administrator, engineer or technician; medium income; no education/primary school/secondary/high school/professional training; and working in the informal sector. In contrast, people who were male, healthcare personnel, high-income earners, participants who do not consume traditional herbal remedies, infected or knowing someone who has been infected by COVID-19, and having a chronic illness or comorbidity, were associated with COVID-19 vaccine acceptance. Participants also gave several reasons they were either hesitant or willing to take the vaccine. A more rigorous surveillance system is needed to systematically monitor drivers of vaccine hesitancy, establish tailored interventions promoting vaccine acceptance, and evaluate the impact of these interventions.
Assessment of Paratuberculosis Vaccination Effect on In Vitro Formation of Neutrophil Extracellular Traps in a Sheep Model
Authors: Arteche-Villasol Noive , Gutiérrez-Expósito Daniel , Criado Miguel , Benavides Julio , Pérez Valentín
Published: August 2022

Vaccination of domestic ruminants against paratuberculosis has been related to homologous and heterologous protective effects that have been attributed to the establishment of a trained immune response. Recent evidence suggests that neutrophils could play a role in its development. Therefore, we propose an in vitro model for the study of the effect of paratuberculosis vaccination on the release of neutrophil extracellular traps (NETs) in sheep. Ovine neutrophils were obtained from non-vaccinated (n = 5) and vaccinated sheep (n = 5) at different times post-vaccination and infected in vitro with Mycobacterium avium subsp. paratuberculosis (Map), Staphylococcus aureus (SA), and Escherichia coli (EC). NETs release was quantified by fluorimetry and visualized by immunofluorescence microscopy. Typical NETs components (DNA, neutrophil elastase, and myeloperoxidase) were visualized extracellularly in all infected neutrophils; however, no significant percentage of extracellular DNA was detected in Map-infected neutrophils compared with SA- and EC-infected. In addition, no significant effect was detected in relation to paratuberculosis vaccination. Further assays to study NETs release in ovine neutrophils are needed. Preliminary results suggest no implication of NETs formation in the early immune response after vaccination, although other neutrophil functions should be evaluated.
Tyrosine Kinase Inhibitors Do Not Promote a Decrease in SARS-CoV-2 Anti-Spike IgG after BNT162b2 Vaccination in Chronic Myeloid Leukemia: A Prospective Observational Study
Authors: Katagiri Seiichiro , Akahane Daigo , Otsuki Shunsuke , Suto Arisa , Yamada Akiko , Suguro Tamiko , Asano Michiyo , Yoshizawa Seiichiro , Tanaka Yuko , Furuya Nahoko , Fujimoto Hiroaki , Okabe Seiichi , Gotoh Moritaka , Ito Yoshikazu , Gotoh Akihiko
Published: August 2022

We performed a prospective observational study of chronic myeloid leukemia (CML) patients after anti-SARS-CoV-2 BNT162b2 vaccination (VC). In total, 32 CML patients with tyrosine kinase inhibitor (TKI) therapy, 10 CML patients with treatment-free remission, and 16 healthy subjects participated in the study. From April 2021 to September 2021, all cases (median age = 58 years) were vaccinated twice. Immunoglobulin G for SARS-CoV-2 spike protein (S-IgG) was measured at three timepoints (before the first VC, 1–5 weeks after the second VC (T1), and approximately 6 months after the second VC (T2)). S-IgG was not observed before the first VC in any participant. At T1, all cases had acquired S-IgG. There were no significant differences in S-IgG levels among groups. A paired sample comparison of median S-IgG titers between T1 and T2 in all groups showed a significant reduction in T2 S-IgG titers. There were no significant differences in S-IgG levels among groups. When all patients were analyzed, those aged ≥58 years had significantly lower S-IgG levels than those aged <58 years at T1. The BNT162b2 vaccine was highly effective in CML patients with or without TKIs, and S-IgG levels were as persistent as those in healthy individuals.
Clinical Progression and Outcome of Hospitalized Patients Infected with SARS-CoV-2 Omicron Variant in Shanghai, China
Authors: Shao Jiasheng , Fan Rong , Hu Jianrong , Zhang Tiejun , Lee Catherine , Huang Xuyuan , Wang Fei , Liang Haiying , Jin Ye , Jiang Ying , Gu Yanhua , Huang Gang
Published: August 2022

Background: Studies on the Omicron variant infection have generally been restricted to descriptions of its initial clinical and epidemiological characteristics. We investigated the timeline-related progression and clinical outcome in hospitalized individuals with the Omicron variant. Methods: We conducted a retrospective, single-centered study including 226 laboratory-confirmed cases with the Omicron variant between 6 April and 11 May 2022 in Shanghai, China. The final date of follow-up was 30 May 2022. Results: Among 226 enrolled patients, the median age was 52 years, and 118 (52.2%) were female. The duration from onset of symptoms to hospitalization was 3 days (interquartile range (IQR): 2–4 days) for symptomatic patients. Cough occurred in 168 patients (74.3%). The median interval to negative reverse-transcriptase PCR tests of nasopharynx swab was 10 days ((IQR): 8–13 days). No radiographic progressions were found in 196 patients on the 7th day after onset of symptoms. The median duration of fever in all participants was 5 days (IQR: 4–6 days). The median PCR conversion time of Paxlovid-treated patients was 8 days (IQR: 7–10 days) compared with that of a traditional Chinese herb medicine lianhuaqingwen (10 days, IQR: 8–13 days) (p = 0.00056). Booster vaccination can significantly decrease the severity of Omicron infection when compared with unvaccinated patients (p = 0.009). In multivariate logistic analysis, erythrocyte sedimentation rate (ESR) (OR = 1.05) was independently related to the severity of the infection. Conclusions: The majority of clinical symptoms of Omicron infection were not severe. Early and aggressive administration of Paxlovid can significantly reduce the PCR conversion time. Booster vaccination should also be highly recommended in the population over 14 years old.
Japanese Encephalitis DNA Vaccines with Epitope Modification Reduce the Induction of Cross-Reactive Antibodies against Dengue Virus and Antibody-Dependent Enhancement of Dengue Virus Infection
Authors: Kotaki Tomohiro , Nagai Yurie , Yamanaka Atsushi , Konishi Eiji , Kameoka Masanori
Published: August 2022

Infection with viruses belonging to the genus Flavivirus, such as Japanese encephalitis virus (JEV) and dengue virus (DENV), is a worldwide health problem. Vaccines against JEV and DENV are currently available. However, the dengue vaccine possibly increases the risk of severe dengue due to antibody-dependent enhancement (ADE). Moreover, the Japanese encephalitis (JE) vaccine reportedly induces cross-reactive ADE-prone antibodies against DENV, potentially leading to symptomatic dengue. Therefore, it is necessary to eliminate the risk of ADE through vaccination. In this study, we attempted to develop a JE vaccine that does not induce ADE of DENV infection using an epitope modification strategy. We found that an ADE-prone monoclonal antibody cross-reactive to DENV and JEV recognizes the 106th amino acid residue of the E protein of JEV (E-106). The JE DNA vaccine with a mutation at E-106 (E-106 vaccine) induced comparable neutralizing antibody titers against JEV to those induced by the wild-type JE DNA vaccine. Meanwhile, the E-106 vaccine induced 64-fold less cross-reactive ADE-prone antibodies against DENV. The mutation did not compromise the protective efficacy of the vaccine in the lethal JEV challenge experiment. Altogether, the modification of a single amino acid residue identified in this study helped in the development of an ADE-free JE vaccine.
Phase I Study to Assess the Safety and Immunogenicity of an Intradermal COVID-19 DNA Vaccine Administered Using a Pyro-Drive Jet Injector in Healthy Adults
Authors: Nakagami Hironori , Hayashi Hiroki , Sun Jiao , Yanagida Yuka , Otera Takako , Nakagami Futoshi , Hamaguchi Shigeto , Yoshida Hisao , Okuno Hideo , Yoshida Shota , Nakamaru Ryo , Yokoyama Serina , Fujimoto Taku , Hongyo Kazuhiro , Akeda Yukihiro , Morishita Ryuichi , Tomono Kazunori , Rakugi Hiromi
Published: August 2022

We conducted a nonrandomized, open-label phase I study to assess the safety and immunogenicity of an intradermal coronavirus disease 2019 (COVID-19) DNA vaccine (AG0302-COVID-19) administered using a pyro-drive jet injector at Osaka University Hospital between Yanagida November 2020 and December 2021. Twenty healthy volunteers, male or female, were enrolled in the low-dose (0.2 mg) or high-dose (0.4 mg) groups and administered AG0302-COVID19 twice at a 2-week interval. There were no adverse events that led to discontinuation of the study drug vaccination schedule. A serious adverse event (disc protrusion) was reported in one patient in the high-dose group, but the individual recovered, and the adverse event was not causally related to the study drug. In the analysis of the humoral immune response, the geometric mean titer (GMT) of serum anti-SARS-CoV-2 spike glycoprotein-specific antibody was low in both the low-dose and high-dose groups (246.2 (95% CI 176.2 to 344.1, 348.2 (95% CI 181.3 to 668.9)) at the 8 weeks after first vaccination. Regarding the analysis of the cellular immune, the number of IFN-γ-producing cells responsive to the SARS-CoV-2 spike glycoprotein increased with individual differences after the first dose and was sustained for several months. Overall, no notable safety issues were observed with the intradermal inoculations of AG0302-COVID19. Regarding immunogenicity, a cellular immune response was observed in some subjects after AG0302-COVID19 intradermal inoculation, but no significant antibody production was observed.