Genetic analysis of porcine H3N2 viruses originating in southern China

From immunological and phylogenetic analyses of H3 influenza viruses isolated from pigs and ducks in the People's Republic of China (China), Hong Kong, Taiwan and Japan, between 1968 and 1982, we arrived at the following conclusions. The H3 haemagglutinin and N2 neuraminidase genes from swine isolates can be segregated into four mammalian lineages, including: (i) the earliest human strains; (ii) early swine strains including Hong Kong isolates from 1976-1977; (iii) an intermediate strain between the early swine and recent human strains; and (iv) recent human strains. In this study we found an unusual swine strain (sw/Hong Kong/127/82) belonging to the third lineage which behaved like those of the early swine-like lineage in the haemagglutination inhibition test; but neuraminidase inhibition profiles with monoclonal antibodies indicated that this virus is related to late human strains. On the basis of pairwise comparisons of complete or partial nucleotide sequences the genes encoding the three polymerase proteins (PB2, PB1, PA), the nucleoprotein, the membrane protein and possibly the nonstructural proteins of sw/Hong Kong/127/82 are of the swine H1N1 lineage, whereas genes encoding the two surface glycoproteins belong to the human H3N2 lineage. In contrast, all RNA segments of one swine isolate (sw/Hong Kong/81/78) are similar to those of recent human H3N2 viruses. This study indicated that frequent interspecies infections between human and swine hosts appeared to occur during 1976-82. Although the evolutionary rates of human (0.0122/site/year), swine (0.0127/site/year) and avian (0.0193/site/year) virus genes are similar when based upon synonymous substitutions, nonsynonymous substitutions indicated that viral genes derived from human and swine viruses evolved about three times faster (0.0026-0.0027/site/year) than those of avian viruses (0.0008/site/year). Furthermore, the evolutionary mechanism by which human and swine H3 haemagglutinin genes evolve at a similar rate, based on nonsynonymous substitutions, appeared to be quite different from previous evidence which showed that human H1 haemagglutinin genes evolved three times faster than those of swine viruses. However, comparison of the number of nonsynonymous substitutions in the antigenic sites (A-E) of haemagglutinin molecules demonstrated that swine viruses evolve at a rate that is about one fifth to one tenth that of human viruses, reflecting the conservative nature of the antigenic structure in the former.published_or_final_versio

Workshop 1: Surveillance issues of pandemic influenza

published_or_final_versio

Potent and broad anticancer activities of leaf extracts from Melia azedarach L. of the subtropical Okinawa islands

Plant extracts have been traditionally used for various therapeutic applications. By conducting an initial screening of several subtropical plants, in this study, we evaluated the anticancer activities of Melia azedarach L. The extract from Melia azedarach L. leaves (MLE) show high cytotoxic effects on cancer cells and in vivo mouse and dog tumor models. During the initial screening, MLE showed strong antiproliferative activity against HT-29 colon, A549 lung, and MKN1 gastric cancer cells. In subsequent tests, using 39 human tumor cell lines, we confirmed the potent anticancer activities of MLE. The anticancer activity of MLE was also confirmed in vivo. MLE markedly inhibited the growth of transplanted gastric MKN1 cancer xenografts in mice. To elucidate the mechanism underlying the anticancer effects of MLE, MLE-treated MKN1 cells were observed using an electron microscope; MLE treatment induced autophagy. Furthermore, western blot analysis of proteins in lysates of MLE-treated cells revealed induction of light chain 3 (LC3)-II autophagosomal proteins. Thus, MLE appeared to suppress MKN1 cell proliferation by inducing autophagy. In addition, in the mouse macrophage cell line J774A.1, MLE treatment induced TNF-alpha production, which might play a role in tumor growth suppression in vivo. We also performed a preclinical evaluation of MLE treatment on dogs with various cancers in veterinary hospitals. Dogs with various types of cancers showed a mean recovery of 76% when treated with MLE. Finally, we tried to identify the active substances present in MLE. All the active fractions obtained by reverse-phase chromatography contained azedarachin B-related moieties, such as 3-deacetyl-12-hydroxy-amoorastatin, 12-hydroxy-amoorastatin, and 12-hydroxyamoorastaton. In conclusion, MLE contains substances with promising anticancer effects, suggesting their future use as safe and effective anticancer agents

Functional Ingredients Extraction from Garcinia mangostana Pericarp by Liquefied Dimethyl Ether

The mangosteen (Garcinia mangostana Linn.) pericarp contains rich xanthone, a one kind of the polyphenols in the non-edible portion. In recent years, xanthones have been noted as a functionality such as anti-cancer effect and is expected as pharmaceuticals and health supplements. In this work, extraction of xanthones from mangosteen pericarp by using liquefied DME were investigated. Wet powder and cube samples were used as a raw material. Experimental conditions were 35°C, 0.8 MPa with various amounts of sample (1, 3, 6 g). Extracted components were analyzed by using high performance liquid chromatography (HPLC). As a result, it was confirmed that eight kinds of xanthones such as alpha-mangostin, 3-Isomangostin, Mangostanol, 8-Desoxygartanin, Gartanin, Garcinone E, 9-Hydroxycalabaxanthone, beta-Mangostin were extracted. The highest yield of alpha-mangostin (42.9 mg/g_dry sample) was obtained with a powder sample of 6 g loaded. Final alpha-magostin corresponded to about 104% of the ethanol extraction with wet mangosteen pericarp, and 72% of the dried sample. Therefore, it was considered that in the extraction of xanthones from the mangosteen, liquefied DME extraction was valid

Initial In-Orbit Operation Result of Microsatellite HIBARI: Attitude Control by Driving Solar Array Paddles

We have developed a 50kg class microsatellite HIBARI . The mission of this satellite is to demonstrate a novel attitude control method for microsatellites which is called “Variable Shape Attitude Control (VSAC).” VSAC is a method using anti-torque by driving variable shape structures. HIBARI has four drivable solar array paddles, and will demonstrate VSAC. The development of HIBARI began in 2019, and it was injected into orbit in November 2021 under the Innovative Satellite Technology Demonstration Program led by JAXA. Currently, HIBARI has completed its critical phase and paddle deployment phase, and is conducting paddle drive experiments in orbit. In paddle drive experiments, the paddles are driven according to the command values, and the accompanying attitude change is confirmed. These results indicate that the satellite can generate angular velocities of 4 deg/s or more and achieve the target agile maneuver of 30deg in 10seconds, which is comparable to that of CMG for microsatellite

Daily mobility and centrality restructuring in the south of the metropolitan area of Buenos Aires

Desde inicios de los 2000, se registran en la Región Metropolitana de Buenos Aires procesos de densificación y renovación de algunos subcentros metropolitanos, estructurados a lo largo de antiguas arterias y ejes ferroviarios, con atributos de compacidad y continuidad con el tejido existente, y con un radio de influencia que supera el del municipio de pertenencia. Pese a no haber cesado el crecimiento residencial disperso y la profusión de enclaves comerciales y de servicios asociados a un patrón de movilidad “automóvil intensivo”, se considera necesario atender estas otras dinámicas, incluyendo la revitalización del comercio de calle y la movilidad por transporte público, como modelos alternativos de reestructuración de la centralidad metropolitana. Dentro de este marco, el artículo analiza los patrones de movilidad cotidiana y los procesos de verticalización residencial, localización de franquicias y sus sinergias con otras actividades comerciales y de servicio en tres subcentros del corredor sur (Avellaneda, Lanús y Lomas de Zamora). Junto al mapeo e interpretación de los relevamientos, se exponen los principales resultados de las encuestas de movilidad, evidenciando las interacciones que existen entre los centros de estudio, así como la atracción que ejercen respecto de sus municipios vecinos y su potencial capacidad de desarrollo como centralidades de referencia a escala regional. Since the beginning of 2000, the Buenos Aires Metropolitan Region has registered processes of renovation and densification of some of its metropolitan sub-centres, structured along old arteries and railways routes, with attributes of compactness and continuity with the existing urban fabric and also, with a radius of influence that exceeds the municipality to which they belong. Despite the growth of scattered housing and the profusion of commercial and services enclaves related with an “automobile-intensive” mobility pattern, it is considered necessary to address these other dynamics, including the revitalization of the street commerce and the public transportation mobility, as alternative models of restructuration for the metropolitan centrality. Within this framework, the article analyzes daily mobility patterns, processes of residential verticalization, location of franchises and their synergies with other commercial and services activities in three sub-centres of the southern corridor: Avellaneda, Lanús and Lomas de Zamora. Along with the mapping and interpretation of data collection, the article describes the main results of mobility surveys, showing the interactions between the centers studied as well as the attraction they exert regarding their neighboring municipalities and its potential ability to develop as regional reference centers.

Engineering Model Development of HIBARI: MicroSatellite for Technology Demonstration of Variable-Shape Attitude Control

We are developing a 40kg class microsatellite “HIBARI”. The main technical mission is demonstration a novel attitude control method called “Variable Shape Attitude Control (VSAC)” proposed by Matunaga, Tokyo Institute of Technology. This VSAC is based on an idea to utilize a reaction torque generated by changing the shape of satellites, for example driving solar array paddles by actuators. HIBARI is planned to be launched in fiscal year 2021 under “Innovative Satellite Technology Demonstration Program” led by JAXA. We are developing EM of HIBARI and describes those in this paper. Specifically, the results of missions, systems, and various tests are shown and the validity is derived

Integrating influenza antigenic dynamics with molecular evolution.

Influenza viruses undergo continual antigenic evolution allowing mutant viruses to evade host immunity acquired to previous virus strains. Antigenic phenotype is often assessed through pairwise measurement of cross-reactivity between influenza strains using the hemagglutination inhibition (HI) assay. Here, we extend previous approaches to antigenic cartography, and simultaneously characterize antigenic and genetic evolution by modeling the diffusion of antigenic phenotype over a shared virus phylogeny. Using HI data from influenza lineages A/H3N2, A/H1N1, B/Victoria and B/Yamagata, we determine patterns of antigenic drift across viral lineages, showing that A/H3N2 evolves faster and in a more punctuated fashion than other influenza lineages. We also show that year-to-year antigenic drift appears to drive incidence patterns within each influenza lineage. This work makes possible substantial future advances in investigating the dynamics of influenza and other antigenically-variable pathogens by providing a model that intimately combines molecular and antigenic evolution. DOI: http://dx.doi.org/10.7554/eLife.01914.001

Mammalian innate resistance to highly pathogenic avian influenza H5N1 virus infection is mediated through reduced proinflammation and infectious virus release

Respiratory epithelial cells and macrophages are the key innate immune cells that play an important role in the pathogenesis of influenza A virus infection. We found that these two cell types from both human and pig showed comparable susceptibilities to initial infection with a highly pathogenic avian influenza (HPAI) H5N1 virus (A/turkey/Turkey/1/05) and a moderately pathogenic human influenza H1N1 virus (A/USSR/77), but there were contrasting differences in host innate immune responses. Human cells mounted vigorous cytokine (tumor necrosis factor alpha [TNF-α] and interleukin-6 [IL-6]) and chemokine (CXCL9, CXCL10, and CXCL11) responses to H5N1 virus infection. However, pig epithelial cells and macrophages showed weak or no TNF-α and chemokine induction with the same infections. The apparent lack of a strong proinflammatory response, corroborated by the absence of TNF-α induction in H5N1 virus-challenged pigs, coincided with greater cell death and the reduced release of infectious virus from infected pig epithelial cells. Suppressor of cytokine signaling 3 (SOCS3), a protein suppressor of the JAK-STAT pathway, was constitutively highly expressed and transcriptionally upregulated in H5N1 virus-infected pig epithelial cells and macrophages, in contrast to the corresponding human cells. The overexpression of SOCS3 in infected human macrophages dampened TNF-α induction. In summary, we found that the reported low susceptibility of pigs to contemporary Eurasian HPAI H5N1 virus infections coincides at the level of innate immunity of respiratory epithelial cells and macrophages with a reduced output of viable virus and an attenuated proinflammatory response, possibly mediated in part by SOCS3, which could serve as a target in the treatment or prevention of virus-induced hypercytokinemia, as observed for humans

Novel avian paramyxovirus isolated from gulls in Caspian seashore in Kazakhstan.

Three isolates APMV/gull/Kazakhstan/5976/2014, APMV/gull/Kazakhstan/ 5977/2014 and APMV/gull/Kazakhstan/5979/2014, were obtained from independent samples during annual surveillance for avian influenza and paramyxoviruses in wild birds from the Caspian Sea coast in Western Kazakhstan, and were initially identified as putative paramyxoviruses on the basis of electron microscopy. Hemagglutination Inhibition Assays with antisera to nine known APMV serotypes (APMV1-9) indicated no relation to any of them. Next generation sequencing of whole genome sequences indicated the three isolates were genetically identical, and had a nucleotide structure typical for all APMVs, consisting of six genes 3'-NP-P-M-F-HN-L-5'. Phylogenetic analyses, and assessment of amino acid identities, suggested the most closely related lineages to be APMV-2, 8, 10 and 15, but the novel isolate had less than 64% identity to them and all other known avian paramyxoviruses. This value was above levels considered to generally define other APMV serotypes. Estimates of the evolutionary divergence of the nucleotide sequences of the genomes of APMVs have shown that novel Kazakhstan APMV strain was closest to APMV-2, APMV-8, APMV-10 and APMV-15, with calculated distance values of 2.057, 2.058, 2.026 and 2.286 respectively, which is above values considered to differentiate other serotypes (observed minimum was 1.108 between APMV-1 and recently isolated APMV/UPO216/Korea). Together, the data suggest that isolate APMV/gull/Kazakhstan/5976/2014 and other two should be considered as the first representative of a novel APMV-20 group, and is the first time that avian paramyxoviruses have been found infecting members of the gull family, extending the known taxonomic host range