Isolation and characterization of mammalian orthoreovirus from bats in the United States.

Mammalian orthoreovirus (MRV) infects many mammalian species including humans, bats, and domestic animals. To determine the prevalence of MRV in bats in the United States, we screened more than 900 bats of different species collected during 2015-2019 by a real-time reverse-transcription polymerase chain reaction assay; 4.4% bats tested MRV-positive and 13 MRVs were isolated. Sequence and phylogenetic analysis revealed that these isolates belonged to four different strains/genotypes of viruses in Serotypes 1 or 2, which contain genes similar to those of MRVs detected in humans, bats, bovine, and deer. Further characterization showed that these four MRV strains replicated efficiently on human, canine, monkey, ferret, and swine cell lines. The 40/Bat/USA/2018 strain belonging to the Serotype 1 demonstrated the ability to infect and transmit in pigs without prior adaptation. Taken together, this is evidence for different genotypes and serotypes of MRVs circulating in US bats, which can be a mixing vessel of MRVs that may spread to other species, including humans, resulting in cross-species infections.

Pathogenicity of modified bat influenza virus with different M genes and its reassortment potential with swine influenza A virus.

In our previous studies, the reassortant virus containing only the PR8 H1N1 matrix (M) gene in the background of the modified bat influenza Bat09 : mH1mN1 virus could be generated. However, whether M genes from other origins can be rescued in the background of the Bat09 : mH1mN1 virus and whether the resulting novel reassortant virus is virulent remain unknown. Herein, two reassortant viruses were generated in the background of the Bat09 : mH1mN1 virus containing either a North American or a Eurasian swine influenza virus M gene. These two reassortant viruses and the reassortant virus with PR8 M as well as the control Bat09 : mH1mN1 virus replicated efficiently in cultured cells, while the reassortant virus with PR8 M grew to a higher titre than the other three viruses in tested cells. Mouse studies showed that reassortant viruses with either North American or Eurasian swine influenza virus M gene did not enhance virulence, whereas the reassortant virus with PR8 M gene displayed higher pathogenicity when compared to the Bat09 : mH1mN1 virus. This is most likely due to the fact that the PR8 H1N1 virus is a mouse-adapted virus. Furthermore, reassortment potential between the Bat09 : mH1mN1 virus and an H3N2 swine influenza virus (A/swine/Texas/4199-2/1998) was investigated using co-infection of Madin-Darby canine kidney cells, but no reassortant viruses were detected. Taken together, our results indicate that the modified bat influenza virus is most likely incapable of reassortment with influenza A viruses with in vitro co-infection experiments, although reassortant viruses with different M genes can be generated by reverse genetics.

Effects of PB1-F2 on the pathogenicity of H1N1 swine influenza virus in mice and pigs.

Although several studies have exploited the effects of PB1-F2 in swine influenza viruses, its contribution to the pathogenicity of swine influenza viruses remains unclear. Herein, we investigated the effects of PB1-F2 on the pathogenicity of influenza virus using a virulent H1N1 A/swine/Kansas/77778/2007 (KS07) virus, which expresses a full-length PB1-F2, in mice and pigs. Using reverse genetics, we generated the wild-type KS07 (KS07_WT), a PB1-F2 knockout mutant (KS07_K/O) and its N66S variant (KS07_N66S). KS07_K/O showed similar pathogenicity in mice to the KS07_WT, whereas KS07_N66S displayed enhanced virulence when compared to the other two viruses. KS07_WT exhibited more efficient replication in lungs and nasal shedding in infected pigs than the other two viruses. Pigs infected with the KS07_WT had higher pulmonary levels of granulocyte-macrophage colony-stimulating factor, IFN-γ, IL-6 and IL-8 at 3 and 5 days post-infection, as well as lower levels of IL-2, IL-4 and IL-12 at 1 day post-infection compared to those infected with the KS07_K/O. These results indicate that PB1-F2 modulates KS07 H1N1 virus replication, pathogenicity and innate immune responses in pigs and the single substitution at position 66 (N/S) in the PB1-F2 plays a critical role in virulence in mice. Taken together, our results provide new insights into the effects of PB1-F2 on the virulence of influenza virus in swine and support PB1-F2 as a virulence factor of influenza A virus in a strain- and host-dependent manner.

Pathogenicity and transmissibility of novel reassortant H3N2 influenza viruses with 2009 pandemic H1N1 genes in pigs.

UNLABELLED: At least 10 different genotypes of novel reassortant H3N2 influenza viruses with 2009 pandemic H1N1 [A(H1N1)pdm09] gene(s) have been identified in U.S. pigs, including the H3N2 variant with a single A(H1N1)pdm09 M gene, which has infected more than 300 people. To date, only three genotypes of these viruses have been evaluated in animal models, and the pathogenicity and transmissibility of the other seven genotype viruses remain unknown. Here, we show that three H3N2 reassortant viruses that contain 3 (NP, M, and NS) or 5 (PA, PB2, NP, M, and NS) genes from A(H1N1)pdm09 were pathogenic in pigs, similar to the endemic H3N2 swine virus. However, the reassortant H3N2 virus with 3 A(H1N1)pdm09 genes and a recent human influenza virus N2 gene was transmitted most efficiently among pigs, whereas the reassortant H3N2 virus with 5 A(H1N1)pdm09 genes was transmitted less efficiently than the endemic H3N2 virus. Interestingly, the polymerase complex of reassortant H3N2 virus with 5 A(H1N1)pdm09 genes showed significantly higher polymerase activity than those of endemic and reassortant H3N2 viruses with 3 A(H1N1)pdm09 genes. Further studies showed that an avian-like glycine at position 228 at the hemagglutinin (HA) receptor binding site is responsible for inefficient transmission of the reassortant H3N2 virus with 5 A(H1N1)pdm09 genes. Taken together, our results provide insights into the pathogenicity and transmissibility of novel reassortant H3N2 viruses in pigs and suggest that a mammalian-like serine at position 228 in the HA is critical for the transmissibility of these reassortant H3N2 viruses. IMPORTANCE: Swine influenza is a highly contagious zoonotic disease that threatens animal and public health. Introduction of 2009 pandemic H1N1 virus [A(H1N1)pdm09] into swine herds has resulted in novel reassortant influenza viruses in swine, including H3N2 and H1N2 variants that have caused human infections in the United States. We showed that reassortant H3N2 influenza viruses with 3 or 5 genes from A(H1N1)pdm09 isolated from diseased pigs are pathogenic and transmissible in pigs, but the reassortant H3N2 virus with 5 A(H1N1)pdm09 genes displayed less efficient transmissibility than the endemic and reassortant H3N2 viruses with 3 A(H1N1)pdm09 genes. Further studies revealed that an avian-like glycine at the HA 228 receptor binding site of the reassortant H3N2 virus with 5 A(H1N1)pdm09 genes is responsible for less efficient transmissibility in pigs. Our results provide insights into viral pathogenesis and the transmission of novel reassortant H3N2 viruses that are circulating in U.S. swine herds and warrant future surveillance.

Characterization of uncultivable bat influenza virus using a replicative synthetic virus.

Bats harbor many viruses, which are periodically transmitted to humans resulting in outbreaks of disease (e.g., Ebola, SARS-CoV). Recently, influenza virus-like sequences were identified in bats; however, the viruses could not be cultured. This discovery aroused great interest in understanding the evolutionary history and pandemic potential of bat-influenza. Using synthetic genomics, we were unable to rescue the wild type bat virus, but could rescue a modified bat-influenza virus that had the HA and NA coding regions replaced with those of A/PR/8/1934 (H1N1). This modified bat-influenza virus replicated efficiently in vitro and in mice, resulting in severe disease. Additional studies using a bat-influenza virus that had the HA and NA of A/swine/Texas/4199-2/1998 (H3N2) showed that the PR8 HA and NA contributed to the pathogenicity in mice. Unlike other influenza viruses, engineering truncations hypothesized to reduce interferon antagonism into the NS1 protein didn't attenuate bat-influenza. In contrast, substitution of a putative virulence mutation from the bat-influenza PB2 significantly attenuated the virus in mice and introduction of a putative virulence mutation increased its pathogenicity. Mini-genome replication studies and virus reassortment experiments demonstrated that bat-influenza has very limited genetic and protein compatibility with Type A or Type B influenza viruses, yet it readily reassorts with another divergent bat-influenza virus, suggesting that the bat-influenza lineage may represent a new Genus/Species within the Orthomyxoviridae family. Collectively, our data indicate that the bat-influenza viruses recently identified are authentic viruses that pose little, if any, pandemic threat to humans; however, they provide new insights into the evolution and basic biology of influenza viruses.

Initial phantom study comparing image quality in computed tomography using adaptive statistical iterative reconstruction and new adaptive statistical iterative reconstruction v.

PURPOSE: The purpose of this study was to assess the image quality of a novel advanced iterative reconstruction (IR) method called as "adaptive statistical IR V" (ASIR-V) by comparing the image noise, contrast-to-noise ratio (CNR), and spatial resolution from those of filtered back projection (FBP) and adaptive statistical IR (ASIR) on computed tomography (CT) phantom image. MATERIALS AND METHODS: We performed CT scans at 5 different tube currents (50, 70, 100, 150, and 200 mA) using 3 types of CT phantoms. Scanned images were subsequently reconstructed in 7 different scan settings, such as FBP, and 3 levels of ASIR and ASIR-V (30%, 50%, and 70%). The image noise was measured in the first study using body phantom. The CNR was measured in the second study using contrast phantom and the spatial resolutions were measured in the third study using a high-resolution phantom. We compared the image noise, CNR, and spatial resolution among the 7 reconstructed image scan settings to determine whether noise reduction, high CNR, and high spatial resolution could be achieved at ASIR-V. RESULTS: At quantitative analysis of the first and second studies, it showed that the images reconstructed using ASIR-V had reduced image noise and improved CNR compared with those of FBP and ASIR (P < 0.001). At qualitative analysis of the third study, it also showed that the images reconstructed using ASIR-V had significantly improved spatial resolution than those of FBP and ASIR (P < 0.001). CONCLUSIONS: Our phantom studies showed that ASIR-V provides a significant reduction in image noise and a significant improvement in CNR as well as spatial resolution. Therefore, this technique has the potential to reduce the radiation dose further without compromising image quality.

Three-month treatment response and exacerbation in chronic obstructive pulmonary disease.

The aim of this study was to investigate relationships between acute exacerbation and Forced Expiratory Volume 1 second (FEV1) improvement after treatment with combined long-acting beta-agonist (LABA) and inhaled corticosteroid (ICS) in patients with chronic obstructive pulmonary disease (COPD). A total of 137 COPD patients were classified as responders or nonresponders according to FEV1 improvement after 3 months of LABA/ICS treatment in fourteen referral hospitals in Korea. Exacerbation occurrence in these two subgroups was compared over a period of 1 yr. Eighty of the 137 COPD patients (58.4%) were classified as responders and 57 (41.6%) as nonresponders. Acute exacerbations occurred in 25 patients (31.3%) in the responder group and in 26 patients (45.6%) in the nonresponder group (P=0.086). FEV1 improvement after LABA/ICS treatment was a significant prognostic factor for fewer acute exacerbations in a multivariate Cox proportional hazard model adjusted for age, sex, FEV1, smoking history, 6 min walk distance, body mass index, exacerbation history in the previous year, and dyspnea scale.Three-month treatment response to LABA/ICS might be a prognostic factor for the occurrence of acute exacerbation in COPD patients.

Serendipitous discovery of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine derivatives as novel HIV-1 replication inhibitors.

We identified a novel class of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine compounds as potent HIV-1 replication inhibitors serendipitously during the process of evaluation of triazolothienopyrimidine (TTPM) compounds. Herein, we report synthesis and biological evaluation of 2-((phenylsulfonyl)methyl)-thieno[3,2-d]pyrimidine compounds using a cell-based full replication assay to identify thienopyrimidines 6 and 30, which could be further utilized as viable lead compounds.

Poly-tobacco use and mental health in South Korean adolescents.

INTRODUCTION: With the advent of new tobacco products, poly-tobacco use among adolescents is increasing. Smoking among adolescents negatively impacts both their physical and mental health. This study aimed to determine poly-tobacco use among adolescents in South Korea and to identify the mental health problems caused by single-, dual-, and poly-tobacco use. METHODS: Data from 54948 adolescents in the 2020 Korea Youth Behavior Web-based Survey were included. Mental health variables of our primary outcome were loneliness, anxiety, and depression. Descriptive statistics, Rao-Scott χ2 test and complex sample multivariable logistic regression analysis were conducted to determine the association between the type of tobacco product use and mental health. RESULTS: Among the subjects, 95.2% were non-tobacco users, followed by single (3.0%), dual (1.1%), and poly users (0.7%). The subjects with poly-tobacco use had significantly higher rates of loneliness (33.2%, p<0.001), anxiety (22.3%, p<0.001), and depression (49.9%, p<0.001) than those who used fewer tobacco products. Subjects who used poly-tobacco products were 2.13 (95% CI: 1.61-2.83) times more likely to report loneliness, 1.52 (95% CI: 1.12-2.07) times more likely to report anxiety, and 2.18 (95% CI: 1.68-2.82) times more likely to report depression than non-tobacco users. CONCLUSIONS: Among adolescents, poly-tobacco use is associated with symptoms of loneliness, depression, and anxiety, which are internalized mental health problems. Poly-tobacco use warrants early assessment of high-risk groups, education on the need for tobacco-use cessation, and active intervention for the psychological difficulties that these high-risk groups experience.

Nutritional Composition and Safety Aspects of Deep-Sea Whelks (Buccinum tenuissimum Kuroda).

The deep-sea whelk Buccinum tenuissimum Kuroda is highly sought-after as food in East Asian countries, notably, Korea and Japan. However, it lacks official recognition as a food product in Korea. This study aimed to assess its nutritional composition and safety for the potential development of seafood products. The nutritional analysis revealed high protein (13.54-20.47 g/100 g whelk), fat (0.85-8.59 g/100 g whelk), carbohydrate (1.55-12.81 g/100 g whelk), and dietary fiber (1.25-1.95 g/100 g whelk) contents in both muscle and gut samples, with energy contents ranging from 339.11 ± 1.64 to 692.00 ± 3.21 kJ/100 g. Key minerals, including iron, potassium, calcium, and sodium, and essential fatty acids, including eicosapentaenoic acid, docosahexaenoic acid, arachidonic acid, omega-3, and omega-6 fatty acids, were abundant, making it a potential supplementary food. Notably, heavy metal levels met the Korean standards for seafood safety. No trans fats, radioactivity concerning the radioactive isotopes 134Cs/137Cs and 131I, or pathogenic bacteria were detected. This confirms the safety and nutritional value of deep-sea whelks, suggesting their potential for developing seafood products rich in beneficial components, which could enhance nutrition and food security while contributing to economic growth.

Evaluating the Efficiency of Black Soldier Fly (Hermetia illucens) Larvae in Converting Mackerel Head Waste into Valuable Resources.

The seafood processing industry generates significant waste, including mackerel heads (MH), constituting 20-32% of total waste. This study explored the potential of utilizing MH as a feed source for black soldier fly larvae (BSF larvae). BSF larvae are known for their ability to efficiently convert organic materials into nutrient-rich biomass. Five concentrations of MH (0, 10, 20, 30, 40, and 50% in chicken feed) were fed to BSF larvae for eight days. After harvesting, their growth, MH conversion efficiency, nutritional content, and heavy metals reduction potential were measured. BSF larvae showed optimal growth when fed with a feed containing 20% MH, resulting in a 14.36-fold increase in weight compared to the control group, as determined by the Fisher's Least Significant Difference Test. BSF larvae maintained a survival rate of 99.33%. With the lowest feed conversion ratio (FCR) of 2.09 at 20% MH, feed efficiency was improved by up to 65.15%, and feed reduction up to 73.53%. MH enhanced lipid and protein content in BSF larvae. Furthermore, BSF larvae in this study showed higher polyunsaturated fatty acids (PUFA), including eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), as well as other amino acids which are required for breeding animals. The current study highlights the potential of MH as a feed source for BSF larvae, improving nutritional biomass. It also suggests BSF larvae as an eco-friendly option for handling seafood processing waste and as an alternative feed source for animals.

Identification of a series of 1,3,4-trisubstituted pyrazoles as novel hepatitis C virus entry inhibitors.

In this report we describe the identification of novel pyrazole analogs as potent hepatitis C virus (HCV) entry inhibitor. The pyrazoles were identified by our phenotypic high-throughput screening using infectious HCV. A series of pyrazole derivatives was synthesized and evaluated for inhibitory activity against HCV in the infectious cell culture system. Through evaluation of selected compounds we observed that the pyrazoles did not interfere with HCV RNA replication but with viral entry as shown by experiments with HCV replicons and HCV pseudo particles, respectively.

Synthesis and biological evaluation of triazolothienopyrimidine derivatives as novel HIV-1 replication inhibitors.

We identified a novel class of triazolothienopyrimidine (TTPM) compounds as potent HIV-1 replication inhibitors during a high-throughput screening campaign that evaluated more than 200,000 compounds using a cell-based full replication assay. Herein, we report the optimization of the antiviral activity in a cell-based assay system leading to the discovery of aryl-substituted TTPM derivatives (38, 44, and 45), which exhibited significant inhibition of HIV-1 replication with acceptable safety margins. These novel and potent TTPMs could serve as leads for further development.

Cyclophilin A as a New Therapeutic Target for Hepatitis C Virus-induced Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) related to hepatitis B virus (HBV) and hepatitis C virus (HCV) infections is thought to account for more than 80% of primary liver cancers. Both HBV and HCV can establish chronic liver inflammatory infections, altering hepatocyte and liver physiology with potential liver disease progression and HCC development. Cyclophilin A (CypA) has been identified as an essential host factor for the HCV replication by physically interacting with the HCV non structural protein NS5A that in turn interacts with RNA-dependent RNA polymerase NS5B. CypA, a cytosolic binding protein of the immunosuppressive drug cyclosporine A, is overexpressed in many cancer types and often associated with malignant transformation. Therefore, CypA can be a good target for molecular cancer therapy. Because of antiviral activity, the CypA inhibitors have been tested for the treatment of chronic hepatitis C. Nonimmunosuppressive Cyp inhibitors such as NIM811, SCY-635, and Alisporivir have attracted more interests for appropriating CypA for antiviral chemotherapeutic target on HCV infection. This review describes CypA inhibitors as a potential HCC treatment tool that is contrived by their obstructing chronic HCV infection and summarizes roles of CypA in cancer development.

Development of 18F-Labeled PET Tracer Candidates for Imaging of the Abelson Non-receptor Tyrosine Kinase in Parkinson's Disease.

Activated Abelson non-receptor tyrosine kinase (c-Abl) plays a harmful role in neurodegenerative conditions such as Parkinson's disease (PD). Inhibition of c-Abl is reported to have a neuroprotective effect and be a promising therapeutic strategy for PD. We have previously identified a series of benzo[d]thiazole derivatives as selective c-Abl inhibitors from which one compound showed high therapeutic potential. Herein, we report the development of a complementary positron emission tomography (PET) tracer. In total, three PET tracer candidates were developed and eventually radiolabeled with fluorine-18 for in vivo evaluation studies in mice. Candidate [18F]3 was identified as the most promising compound, since it showed sufficient brain uptake, good washout kinetics, and satisfactory metabolic stability. In conclusion, we believe this tracer provides a good starting point to further validate and explore c-Abl as a target for therapeutic strategies against PD supported by PET.

Discovery and characterization of a novel 7-aminopyrazolo[1,5-a]pyrimidine analog as a potent hepatitis C virus inhibitor.

We describe a novel 7-aminopyrazolo[1,5-a]pyrimidine (7-APP) derivative as a potent hepatitis C virus (HCV) inhibitor. A series of 7-APPs was synthesized and evaluated for inhibitory activity against HCV in different cell culture systems. The synthesis and preliminary structure-activity relationship study of 7-APP are reported.

Molecular epidemiologic investigation of lentogenic Newcastle disease virus from domestic birds at live bird markets in Korea.

A Newcastle disease surveillance program was conducted at live bird markets in Korea to expand our epidemiologic understanding of the disease in Korea. During the surveillance program, 10 lentogenic Newcastle disease viruses (NDVs) were isolated and identified from apparently healthy chickens and ducks at live bird markets. The lentogenic viruses had sequence motifs of either 112GKQGRL117 (n = 8) or 112GRQGRL117 (n = 2) at the F0 cleavage site. Sequencing and phylogenetic analyses of NDV isolates based on the hypervariable region of the F protein revealed two different genotypes: genotypes I (n = 8) and II (n = 2). Genotype I viruses were most closely related to the NDV V4 strain (n = 7) or the NDV Ulster 2C strain (n = 1). In contrast, genotype II viruses clustered with the NDV vaccine strains (LaSota and VG/GA) that are commonly used as live vaccines in Korea. The epidemiologic importance of NDV at live bird markets in Korea is discussed.

Psychological Workplace Violence and Health Outcomes in South Korean Nurses.

BACKGROUND: Workplace violence (WPV) and its health consequences should continue to be investigated to foster a healthy and safe working environment, which may reduce nurse staff turnover and improve nurse staff shortages. This study aimed to address the gap in understanding WPV in non-western nurses by examining the relationship between psychological WPV experience, psychological well-being, subjective job stress, and presenteeism among South Korean nurses. METHODS: This cross-sectional study used data from the fifth Korean Working Conditions Survey (KWCS), and 477 nurses were analyzed. For the analysis, selected variables were extracted from the KWCS through a review of the existing literature. In the analysis, we included psychological WPV experiences, such as verbal abuse, unwanted sexual attention, threats, and/or humiliating behaviors over the past 1 month. We measured health outcomes including nurses' psychological well-being, subjective job stress, and presenteeism. FINDINGS: Psychological WPV within the previous month was experienced by 11.1% of the participants. Experience with verbal abuse, threats, or humiliating behaviors was associated with more job stress, higher presenteeism, and poor psychological well-being. CONCLUSION/APPLICATION TO PRACTICE: Study findings suggest that a comprehensive WPV prevention program accompanied by interventions aiming to reduce job stress and improve the well-being of nurses should be actively implemented. To prevent psychological WPV and improve the health of nurses, evidence-based efforts, such as establishment of WPV prevention procedures and education/training of workers at the national, organizational, and individual levels are needed.

Carbapenem-non-susceptible Acinetobacter baumannii of sequence type 92 or its single-locus variants with a G428T substitution in zone 2 of the rpoB gene.

OBJECTIVES: to investigate the epidemiological traits of carbapenem-non-susceptible Acinetobacter baumannii (CNSAB) and the usefulness of phylogenetic grouping based on partial rpoB gene sequencing in defining the epidemiological traits of CNSAB. METHODS: a total of 547 non-duplicate clinical isolates of Acinetobacter spp. were collected from 19 hospitals in Korea in 2008. Detection of genes encoding OXA carbapenemases and metallo-ß-lactamases was performed by PCR. The epidemiological relationships of the isolates were investigated by multilocus sequence typing and repetitive-sequence-based PCR. The 450 bp sequence (zone 2) of the rpoB gene was amplified and sequenced. RESULTS: molecular characterization of the 272 CNSAB isolates identified five sequence types (STs): ST92, ST75, ST137, ST138 and ST69. The first four of these STs were clustered into clonal complex (CC) 92, sharing alleles at six of seven housekeeping gene loci; ST69 shared alleles at five of seven loci. CNSAB of CC92 carried the bla(OXA-23) gene (n = 169), the bla(OXA-51)-like gene preceded by ISAba1 (n = 89) or both (n = 14). Notably, all CNSAB isolates carried a G428T substitution in zone 2 of the rpoB gene. CONCLUSIONS: CNSAB isolates of CC92 with the G428T substitution in zone 2 of the rpoB gene are disseminated nationwide in Korea. A. baumannii with the single nucleotide substitution may be more likely to acquire carbapenem resistance than are other isolates.

Glucosides with cyclic diarylpolynoid as novel C-aryl glucoside SGLT2 inhibitors.

Novel C-aryl glucoside SGLT2 inhibitors containing cyclic diarylpolynoid motif were designed and synthesized for biological evaluation. Alkylzinc bromides have been efficiently prepared by the direct insertion of zinc metal into alkyl bromides. The organozinc reagents underwent smooth Pd-catalyzed cross-coupling reactions. Subsequent ring closing metathesis using 2nd generation Grubbs catalyst successfully generated novel class of ansa-compounds. These glucosides with cyclic diarylpolynoids demonstrated moderate in vitro inhibitory activity against SGLT2 in this series to date (IC(50)=59.5-103 nM).