Tymoviridae comprises positive-sense RNA viruses, which mainly infect plants. Recently, a few Tymoviridae-like viruses have been found in mosquitoes, which feed on vertebrate sources. We describe a novel Tymoviridae-like virus, putatively named, Guachaca virus (GUAV), isolated from Culex pipiens and Culex quinquefasciatus species of mosquitoes and collected in the rural area of Santa Marta, Colombia. After a cytopathic effect was observed in C6/36 cells, RNA was extracted and processed through the NetoVIR next-generation sequencing protocol, and data were analyzed through the VirMAP pipeline. Molecular and phenotypic characterization of the GUAV was achieved using a 5′/3′ RACE, transmission electron microscopy, amplification in vertebrate cells, and phylogenetic analysis. A cytopathic effect was observed in C6/36 cells three days post-infection. The GUAV genome was successfully assembled, and its polyadenylated 3′ end was corroborated. GUAV shared only 54.9% amino acid identity with its closest relative, Ek Balam virus, and was grouped with the latter and other unclassified insect-associated tymoviruses in a phylogenetic analysis. GUAV is a new member of a family previously described as comprising plant-infecting viruses, which seem to infect and replicate in mosquitoes. The sugar- and blood-feeding behavior of the Culex spp., implies a sustained contact with plants and vertebrates and justifies further studies to unravel the ecological scenario for transmission.The family  Tymoviridae belongs to the order Tymovirales, a diverse group represented by viruses that infect plants. The family comprises three genera, Tymovirus, Marafivirus, and Maculavirus, which share their virus morphology with non-enveloped virions and icosahedral capsids of coat proteins that are approximately 30 nm in diameter [Tymovirus, P16 and P31 in Maculavirus, and P21 in Marafivirus [Tymovirus, poly(A)-tailed genomes are characteristic of the genera Maculavirus and Marafivirus (except for Maize rayado fino virus) [The family diameter . These vdiameter . The ideo virus) .Chrysomelidae, Curculionidae, and Cicadellidae may act as vectors of these viruses [Tymoviridae-like viruses in China, Mexico, and Brazil is of major interest [Virus transmission in plants occurs through mechanical contact, infected plant material, or insect vectors ,5. Insec viruses . While a viruses . The mal viruses , which i viruses ,14,15,16interest ,18,19, jTymoviridae, putatively named the Guachaca virus (GUAV), which was isolated from a mosquito pool of the genus Culex in the Sierra Nevada de Santa Marta (SNSM) and located in the north coast of Colombia, an ecosystem that comprises natural (sylvatic) and intervened  areas.Here, we describe the discovery, using molecular and phenotypic characterization, of a new member of the family 2 (6600 sq mi) and serves as the source of 36 rivers. The range is in the Departments of Magdalena, Cesar, and La Guajira. A pool containing six specimens identified through classic taxonomy as belonging to the genus Culex  was collected on 8 June 2018, using a CO2-baited CDC light trap in a rural setting of Quebrada Valencia  near the populated center of Puerto Nuevo-Guachaca area. The mosquito pool was transported in liquid nitrogen from the field station to the molecular biology laboratory at the Universidad Cooperativa de Colombia, where it was stored at −80 °C until processing.The viral metagenomic study in hematophagous mosquitoes was carried out in the natural ecosystem of the Sierra Nevada de Santa Marta, Santa Marta, Magdalena, Colombia. The SNSM is an isolated mountain range in northern Colombia, which is separate from the Andes range that runs through the north of the country. Reaching an elevation of 5700 m  just 42 km (26 mi) from the Caribbean coast, SNSM is the highest coastal range in the tropics, and one of the highest coastal ranges in the world. SNSM encompasses about 17,000 kmAedes albopictus larvae were cultured in Eagle’s minimal essential medium (MEM), supplemented with 10% and 2% fetal bovine serum (FBS) for growth and maintenance, respectively, and incubated in an atmosphere at 95% relative humidity and 5% CO2 at 28 °C. Vero cells  derived from the kidney of the African green monkey Cercopithecus aethiops were cultured in MEM with 8% and 2% FBS for growing and maintenance, respectively, and incubated in an atmosphere at 95% relative humidity and 5% CO2 at 37 °C.C6/36 cells , derived from whole The mosquito pool was mechanically homogenized for 25 s and 1500 rpm in the BeadBug instrument  with the use of ceramic Magna lyser green beads  in 1.3 mL of Dulbecco’s phosphate-buffered saline (DPBS) supplemented with 10% of FBS and 1% of penicillin/streptomycin. Subsequently, it was centrifuged at 14,000 rpm for 15 min and filtered with the use of a 0.4 µm syringe filter. A 100 aliquot of the supernatant was used for inoculation on C6/36 and Vero cell cultures growing in 24-well plates, and the adhesion phase was carried out for 1 h, at 28 °C and 37 °C, respectively. Subsequently, 800 µL of MEM supplemented with 2% FBS and antibiotics were added, and the cultures were incubated at the appropriate temperature. Negative (mock infection) control was included in each multi-well plate. Supernatant of the virus isolation was collected on the third day post-infection when cytopathic effect was evidenced. Subsequently, a second passage was carried out in the same conditions. Each supernatant was collected, cleared, aliquoted, and stored at −70 °C.Virus stocks were obtained from the inoculation of two million C6/36 cells in T25 flasks with a 1/10 dilution of the supernatant of the second passage virus isolation. The inoculation was carried out with 125 µL of the supernatant and 1125 µL of MEM with 2% SFB and 5% tryptose phosphate during one hour at 28 °C. Then, MEM was added to a final volume of 5 mL, and the cell culture was incubated until the cytopathic effect was observed in more than 50% of the cell monolayers around the first three days post-infection. The presence of the virus was confirmed by real-time RT-PCR using custom-made primers as described below.RNA purification from the supernatant of mosquito pool homogenization was performed using the QIAamp Viral RNA mini kit , according to the manufacturer instructions, followed by RNase-free RQ1 DNase treatment to degrade double- and single-stranded DNA . The RNA extract was stored at −80 °C.The RNA extract was used for whole transcriptome amplification through the WTA2 kit  following the NetoVIR protocol . SubsequPrimer design was carried out for the consensus sequence assembled for the GUAV identified in the present study  using thFor 5′ UTR amplification, the 5′/3′ RACE kit second generation  was used. After cDNA synthesis and poly(A) tailing, PCR amplification steps were performed to the poly(A)-tailed cDNA, using the oligo dT-anchor primer and the designed reverse primers, Tymoviridae-like5UTR-R2 and Tymoviridae-like5UTR-R1 , accordi2, 400 µM of each dNTP, 1.25 U of Platinum Taq DNA polymerase , and 5 µL of the polyadenylated cDNA in a final reaction volume of 25 µL. The thermal profile was as follows: 95°C for 4 min, 40 cycles , and a final extension at 72 °C for 5 min. The purification of PCR amplicons was performed using the QIAquick PCR purification kit . DNA sequencing was performed through the Sanger method . The DNA sequences were edited and assembled into the Geneious software v9.1.8 .To corroborate the presence of the poly(A) tail at the 3′ end of the viral genome previously evidenced by the NGS virus assembly, an assay of cDNA synthesis was carried out with random hexamers or Oligo(dT) primer, followed by PCR amplification with the designed Scheme_20_RIGHT_2 (5′-ttacaaaccaggaacctctgact-3′) and the Oligo(dT) primers. The amplification was performed with final concentration of 250 nM of each primer, 1X PCR buffer, 2 mM MgClConsensus sequences of the viral genome obtained from NGS and that of the 3′ UTR obtained by the RACE approach and direct sequencing were assembled through Geneious v9.1.8 , and motifs, secondary structures, and antigenic domains were predicted and annotated. InterPro server from ELIXIR infrastructure was used for functional domain prediction .C6/36 cells growing in T25 flasks were inoculated with a 1/10 dilution of the viral stock and incubated under standard conditions, as described above. Supernatants were collected at 3-, 6-, 9-, 12-, 15-, 18-, 24-, 36-, 48-, and 72 h post-infection, and triplicates were evaluated using real-time RT-PCR. Cell cultures were examined every day for the presence and magnitude of cytopathic effect during the assay.2. Each cell line was grown in the following specific medium conditions: HEK293 in DMEM 10% FBS, HeLa and A549 in MEM 8% FBS, and U937 in RPMI 5% FBS and stimulated with PMA at final concentration of 25 µM. Confluent monolayers or suspensions were inoculated with 500 μL of a 1/10 dilution of the viral stock, following BSL-3 practices. The adhesion phase was carried out at 37 °C for 1 h and then each flask was rinsed twice with 5 mL of DPBS, and 5 mL of fresh culture medium supplemented with 2% FBS were added. Cell culture supernatants were collected at 5, 7, 9, 11, and 13 days post-infection, aliquoted, and stored at −70 °C for subsequent RNA purification and real-time RT-PCR.To assess the ability of GUAV to infect human cells, HEK293 (human embryonic kidney), HeLa , A549 , and U937 (pro monocyte from histiocytic lymphoma) were cultured in T25 flasks at 37 °C with 5% COTymoviridae-like virus were fixed in 3% glutaraldehyde prepared in phosphate buffer pH 7.2, centrifuged at 3000 rpm for 5 min, resuspended, and washed three times with the phosphate buffer pH 7.2. Subsequently, post-fixation with 1% osmium tetroxide was carried out for one hour, followed by three washes, as described above. The cells were dehydrated by treatment with ethanol solutions in ascending concentrations  for 15 min each, followed by propylene oxide for 20 min. For the pre-imbibition, the cell pellet was treated with a mixture of propylene oxide and resin Epon-Araldite in a 2:1 ratio followed by another 1:1 mixture, for one hour. To complete the imbibition, pure resin was added, and the cell pellet was left at 4 °C. The next day, the resin was replaced and the was transferred to an oven at 65 °C for 24 h to obtain the polymerized block. In an ultramicrotome, semi-fine sections (0.5 microns) and ultra-fine sections (60 nanometers) were obtained. The sections were contrasted with uranyl acetate and lead citrate for observation in an electron microscope EM109 .C6/36 cells infected with the Tymoviridae, followed by a multiple sequence alignment (MSA) at the nucleotide and amino acid levels using MAFFT v.7 [A nucleotide sequence dataset was generated through the identification of the ORF1 in previously released sequences of members of the family AFFT v.7 . AccountAFFT v.7  and the AFFT v.7 . Branch AFFT v.7 .Culex spp. (CIST0019) displayed a cytopathic effect that was characterized by the gradual detachment of the cell monolayer by the third day and expanded to the whole culture by the fifth day post-infection. A second passage of a 1/10 dilution of the cell supernatant in fresh C6/36 cells induced the same effect over the following three days, demonstrating the active replication of a transmissible agent , Rinkaby virus , Culex flavivirus , and Atrato Gouko-like virus . Low nucleotide and amino acid identities of 57.8% and 54.9%, respectively, to Ek Balam virus were observed, supporting its proposal as a novel species within the family Tymoviridae, putatively named the Guachaca virus (GUAV).A total of 92,939 paired-end reads were obtained with a Q-score threshold of ≥30. Viral metagenomic analysis using VirMAP  enabled Tymoviridae, with the conserved methyltransferase, endopeptidase, helicase, and RNA polymerase sequence motifs resembling a replicase complex .For phylogenetic analysis, an MSA of 2042 amino acids from a conserved region of the ORF1, ranging from amino acid positions 704 to 1723, was performed for 56 sequences, which were representative of the different genera within the family afivirus . The genTymoviridae (IAT) lineages; one of them is represented by the proposed virus species infecting Bombyx mori, Andrena haemorrhoa, and Lampyris noctiluca, and the second and third IAT-lineages are represented by viruses infecting mosquitoes of the family Culicidae. The second IAT-lineage with a branch support of 100% comprised several proposed species, including the putative GUAV isolated in 2018 in Colombia from Culex spp. mosquitoes; Ek Balam virus, isolated in 2007 in Mexico from Culex quinquefasciatus; and Mutum virus, isolated in 2018 in Brazil from Mansonia sp. A third IAT-lineage of Culicidae-infecting viruses with a branch support of 100% comprised the unclassified Guadeloupe Culex tymo-like virus, Tarnsjo virus, Tymoviridae sp., Sichuan mosquito tymo-like virus, and Culex pseudovishnui tymo-like virus, detected or isolated from Culex spp., and unclassified mosquitoes from Japan, Guadeloupe, Sweden, and China , consistent with their monophyletic grouping (Marafivirus-IAT_Lineage_3 (0.505)  .The TEM results showed complete icosahedral viral particles with a diameter of 40–50 nm in symmetrical arrangements in the cytoplasm, as well as associated with vesicles that were fused to the cell membrane in the cell periphery of C6/36 cells infected with GUAV 5 days post-infection .While GUAV successfully replicated in C6/36 cells, showing an increasing titer during the first 72 hpi , this viCulex of mosquitoes (Diptera: Culicidae) is of special relevance for public health, with several species serving as vectors for critical human and animal pathogens , SLEV, West Nile virus (WNV), etc.). The anthropophilic behavior of this vector species highlights its proximity with humans which, certainly leads to a sustained exposure to the vector virome and the potential transmission of pathogens.The genus Culex spp. that were pool collected in the Sierra Nevada de Santa Marta, an isolated mountain range in the north of Colombia, the cytopathic effect was observed from 3 dpi, gradually extended to the whole monolayer, and was reproduced after a second passage of a diluted inoculum. This finding suggested the presence of a transmissible agent, which was further studied and proposed as a new virus species.After inoculation of a C6/36 cells culture with a supernatant of Ek Balam virus. The metagenomic approach is powerful for the identification of viral signatures, whose genome coverage depends on the viral abundance in a sample. Our results were successful in obtaining the complete genome with high coverage of a new member of the family Tymoviridae, putatively named the Guachaca virus (GUAV). Similar findings have been previously reported in China, Mexico, and Brazil [Tymoviridae were isolated from Culex spp., suggesting the existence of an unclassified group of virus species in this family.Metagenomic approaches for virus discovery have been refined during the last decade, with the publication of several experimental protocols for virus enrichment and pipelines for taxonomic assignment. In our study, the VirMAP pipeline was successfully applied, allowing the identification of a viral contig that is distantly related to all previously reported viruses, but most closely related to the d Brazil ,18,19, wTymovirus, and the absence of the accessory proteins present in the genera Maculavirus and Marafivirus.The Inference of the genome organization of GUAV demonstrated that this virus differed from the three previously defined genera, with the absence of a tRNA-like structure and a movement protein characteristic of the genus The RACE strategy for the 5′ end characterization of the GUAV genome was unsuccessful, but the genome assembly from the NGS data enabled the prediction of secondary RNA structures in that region, starting with a long, perfectly matching stem-loop, followed by a structure of major complexity. The RACE strategy applied to the 3′ end enabled the corroboration of the presence of a poly(A) tail. It is of interest that a highly structured 3′ end with a long stem-loop and internal bulges was successfully identified upstream of the poly(A) tail. Tymoviruses have explored several solutions for the 3′ end stability and function, including unstructured polyadenylated and non-polyadenylated tails, tRNA-like structures [Tymoviridae is mainly represented by plant-infecting viruses, the phylogenetic analysis presented here and in some previously reported studies [While the family  studies ,18,19, sCulex mosquitoes, which is also supported by previous studies with closely related viruses [With the design of a molecular approach, the productive infection in C6/36 cells was demonstrated as an increase in viral genome abundance, which was inversely proportional to the Ct values that gradually decreased between the first- and third-day post-infection. This evidence of infection in mosquito cells indirectly suggests that GUAV could be infecting natural populations of  viruses ,19,30.Tymoviridae in which peripheral vesicles in chloroplasts and/or mitochondria have been reported [The TEM findings evidenced the presence of virus-like particles in the cytoplasm of C6/36 cells and particles associated with intracellular membranes, as has been evidenced for other members of the family reported .All tested mammalian cells were not permissive for GUAV replication. This finding, and the inferred speciation from a family of plant- and insect-associated viruses suggest that the virus circulates in natural cycles, without published evidence of public health implications.Culex spp. vector competence, and virus adaptation to mammalian cells in controlled experimental conditions would help elucidate the molecular and structural basis for receptor binding, host shift, and viral emergence.As part of their ecological interactions warranting virus transmission, several insect species have been incriminated as mechanical or biological vectors. Several insect-specific viruses have been shown to dominate the insect viromes , consideGuachaca virus (GUAV), was discovered and successfully characterized at molecular and phenotypic levels. This virus was unable to replicate in mammalian cells, which suggests its maintenance as an insect-associated virus. While the GUAV biology and transmission cycle remain unraveled, its natural presence in mosquitoes in rural areas, which are in frequent contact with plants and vertebrates as feeding sources, justifies further studies to unravel the ecological scenario for transmission. Phylogenetic evidence warrants a revision of the current taxonomy of the family Tymoviridae to better represent the recently described insect-associated viruses.A novel virus, putatively named the


Hiroshima Peace Memorial Park is widely known as a universal symbol of peace, but there have not been studies of how people actually experience and interpret it. This article presents a detailed case study of a visit to the memorial by using an innovative methodology based on the use of subjective cameras (subcams). Results show that despite the monolithic idea of peace that the memorial officially represents, it is experienced and interpreted in terms of a constant tension which exposes conflicts in post-war Japan memory politics. The dichotomies of war/peace, death/life, past/future, and old /new emerge as part of the participant’s encounter with different situations during his visit. This is particularly clear where he perceives border zones and points of intersection. The article concludes by interpreting these dichotomies through the notion of themata, as elementary dichotomies that underlie a social debate around a specific topic. Specifically, two themata are proposed: one revolving around the temporal problematisation of the past and the future in the memory politics of the A-Bomb, and the other revolving around the spatial dichotomy between the old and the new underlying Hiroshima’s urban renewal. On 6 August 1945, an American B-29 bomber dropped an atomic bomb on the city Hiroshima. The explosion razed the city to the ground and caused the deaths of around 140,000, as a result of direct and indirect effects.This article presents a detailed case study of experiencing and interpreting the Hiroshima memorial. Based on a previous line of research  became a musealised object, which stands today as a material example of the duality that looms behind the memories of the a-bomb in Hiroshima. Visible from the memorial’s Cenotaph and with a central role in Tange’s design for the HPMP, the skeletal remains of the former Industry Promotion Hall stand in stark contrast to the modern city background. However, in addition to bearing a fragment of a past fraught with death and destruction –which contrasts with Hiroshima’s post-war rebirth– the ruins of the A-Bomb Dome also bear witness to the city’s pre-war times by showing a fragment of “a quintessential sign of Japan’s early-twentieth-century imperial modernity” , which record individuals’ ongoing experience from a first-person perspective, in both video and audio Lahlou, . SubcamsThe fieldwork was conducted on 3 December 2021 on the occasion of a research stay by the first author at Kyushu University, hosted by Prof. Minami Hirofumi and funded by the Japan Society for the Promotion of Science. While watching the subcam video of their visit, the SA was able to comment on the experience by reflecting on his affective engagement with the environment and the meanings and associations afforded by some of its elements. As became evident in our previous fieldwork conducted at the Memorial of the Murdered Jews of Europe in Berlin  the memorial entrance, (b) the Cenotaph and (c) the A-Dome. In subsection (d) we include the analysis of the SA’s visit to the Hiroshima National Peace Memorial Hall for the Atomic Bomb Victims, which took place the following day without the subcam glasses. Figure 1) Entering the memorial: a transition from the mundane to the sacred.“Coming to the park from this way, mmm … it is quiet at first. So… your emotions have time to attune to the site, instead of going directly to the memorial’s centre. In this way you are more in control… you do the visit at your own pace”.When the FA and the SA parted ways upon crossing the bridge, the latter initiated the visit by deciding not to go straight to the memorial’s main entrance –where the Gate of the Peace is. He took, instead, the path along the riverside bordering the memorial  ‘Like a time machine’: between past and future, death and life.While watching the video of himself heading towards the area where the Peace Memorial Museum is located, the SA comments on the sense of anticipation he was feeling at the upcoming appearance of the Cenotaph in the central part of the HPMP. This sense of expectation points to the emergent and flowing nature of atmospheres as a sensory quality of experience. As Sumartojo & Pink  point ouThe video shows how he stands contemplating the Cenotaph with the A-Dome in the background from below the Peace Memorial Museum for almost two minutes  Between the old and the new: the symbolic and material power of the A-Dome.The interview goes on as the subcam video progresses showing images of the SA walking along the Peace Pond in the direction of the Children’s Peace Monument without paying much attention to it. After leaving behind that monument, he makes his way towards the twisted Peace Clock Tower which marks the exact time when the bomb was dropped on August 6, thereby bringing the tension between the present and past –made present through the hands of the clock– to the fore once again. From there, the video shows the SA crossing the Aioi BridgeWatching the sunset light casting on the A-Dome, the SA refers to the sacred atmosphere conveyed by these ruins, thus picking up again the duality between the sacred meaning of the HPMP and the mundane life outside the memorial. To this distinction the SA adds the stark contrast generated by “the building in ruins surrounded by more modern architecture”. This is a contrast –between the old and the new, between the city’s past and its modern present– that, in the words of the SA, makes the A-Dome to “stand out more from its surroundings”. Furthermore, the objectification of the past through the A-Dome’s ruins creates a particular atmosphere that feeds into the SA’s imagination when he begins to talk about the people working in the former Industry Promotion Hall: “The people in the building back in 1945 maybe they were busy going about their business and had no time to scape and then vanished in one second”. As Beckstead  points oYou can see how the windows are all blown out and the iron has been twisted, so you can feel the strong wind that came after the bomb […] It’s like a sculpture shaped by the wind of the A-bomb […] This makes this place to have a very special atmosphereDrawing on Niels Bohr’s observations during his visit to Hamlet’s castle in Kronborg, Denmark, Beckstead  stressesLooking at the A-Dome from far away it becomes a symbol, but if you come closer, it gains a more concrete meaning, it becomes something more fragmented and complex. It is not just one thing, one symbol, but a lot of things […] It acquires a more three-dimensional nature, containing a lot of information: the twisted pillars, the windows blown out, …And from that point, he begins to reflect on the complexity of the building’s entrails:schematisation and pleromatisation, two complementary processes mediating aesthetic perception. According to Valsiner . Through pleromatisation new meanings can be created beyond the categorization function of language symbols, something that might be emerging in the post-visit interview. Thus, the detailed description of the A-dome seems to contribute to the emergence of other meanings beyond its official meaning as symbol of peace. More specifically, in focusing on certain aspects of its materiality, the building gains complexity as an intersection point between the idea of peace –officially ascribed to the memorial – and the sense of destruction conveyed through the twisted pillars, the windows blown out, etc.As we can observe, the evocative power of the ruins invites the SA to engage in a meaning-making process as the interview develops. Guided first by the abstract meaning traditionally ascribed to the A-Dome, as a symbol of peace, a closer look at the building – this time through the recorded images of the subcam video– offers the SA the opportunity to reflect on the richness of its materiality. This case is an illustrative example of Valsiner , schemat4) Afterward: from the weight of the past to the lightness of the future.“Everything was solemn and peaceful, and my steps were heavy. In addition, the entire site was built underground. In my mind’s eye I realised how deep the history of this place is buried. There was a tension in the air that contrasted with the atmosphere outside. Seeing the names of the destroyed towns inscribed on the walls gave me a heavy feeling. I was stuck in the underground experience for the rest of the day, even when I walked out of the venue. It was at this point that my view of the city changed again, and I felt as if the entire city had also become heavier. It was then that the sound of a song on the radio, praying for peace, jumped into my ears. I felt the city being reborn, which lifted my spirit at the end of the visit […] So this is a city full of hope, not only full of sad memories. That is what the park transmits to me” (p. 41).The next day, the FA and the SA went to the Hiroshima National Peace Memorial Hall for the Atomic Bomb Victims , thus making the initial stretch of the visit  to be perceived as a liminal zone between the two. This appears to have called for a meaning-making process whereby the notion of a sacred place, applied to the memorial, was set against the idea of mundane and everyday life of the city. Being by the river also generated a dichotomy between war, as a man-made artificial thing associated to the memorial, and nature, implicitly associated with peace.The Cenotaph: The presence of children contributed to create a happy atmosphere leading the SA to contrast the idea of life, hope and future to the deaths of the past associated with the Cenotaph. The intersection between these dichotomic elements is stressed and objectified through a picture the SA took in front of the monument. This dichotomy is latter recreated through the axial view of the Cenotaph at the foreground (associated with a time near at hand) and the A-Dome (representing a past receding into the background) and expressed through the metaphor of the place as a time machine, thus conveying the sense of intersection between past and present.The A-Dome: The dichotomic experience of time shows again when, once close to the A-Dome, the SA highlights the stark contrast between the old ruins and the modern background of the city. This dichotomy between old and new, is complemented by the notion of sacredness attributed to the A-Dome, thus pointing again to an implicit border between the memorial and its mundane surroundings. Finally, the closer look at the A-Dome contributed to the intersection of two contrasting views on the building seen as symbol of peace and as material evidence of war.The Memorial Hall: The dichotomy between past/future, death/life emerges again with the SA’s physical transition from the inside of the Memorial Hall –situated beneath the ground– to the park ground level, where the sound of a recorded music contributed to generate an uplifting feeling from the previous sense of heaviness experienced down in the Hall. Similar to the experience at the memorial entrance, there is the sense of crossing a border separating two worlds endowed with completely different meanings, although this time the transition goes from inside to outside the memorial.As we can see, these dichotomies emerge as a result of the SA’s meaning-making effort at his encounter with different situations during his visit, particularly those leading him to perceive the site in terms of border zones or points of intersection. However, following De Paola et al. , we shouThese two themata correspond to two dimensions –temporal and spatial– involved in the memory politics around the A-Bomb referred to above Schäfer, . The firCombined together these two themata help us to reflect on the problematisation of time and space in the city of Hiroshima. However, one might ask to what degree the existence of different urban topographies linked to dissonant temporalities, as Yoneyama  puts it,Through this article we aimed to address Tange’s question about what crosses people’s minds when visiting the HPMP. To be sure, a defining aspect of HPMP is its location, close to the explosion’s hypocentre. When a memorial is constructed at the site of the traumatic events it is already affectively charged with the site’s historycosmopolitan mode of remembering, the HPMP is inextricably linked to a transnational memory discourse anchored in a set of iconic artefacts –such as the A-Dome – for remembering the past vis-à-vis potential futures we would like to build or avoid as human beings. The global dissemination of images featuring the mushroom cloud, devastated cities, the nuclear reactor, or the radiation symbol have forever changed the way humanity imagines (and questions) its future. As Jasanoff & Kim (sociotechnical imaginaries, imagination of possible futures –whether desirable horizons worth attaining or grim scenarios to be avoided– is becoming increasingly shaped by scientific and technological advances, such as the discovery of nuclear energy.As a paradigmatic example of what Bull & Hansen  call a cff & Kim  examine Yet, imagination about the future is also largely dependent on how we remember the past, and vice versa Brescó, . Thus, a


The transferred LSMO films exhibit a fourfold magnetic anisotropy with easy axis along LSMO < 110 >. A coexistence of uniaxial and fourfold magnetic anisotropy with LSMO [110] easy axis is observed for the 45° Sample by applying a 7.2 kV cm−1 electrical field, significantly different from a uniaxial anisotropy with LSMO [100] easy axis for the 0° Sample. The fitting of the ferromagnetic resonance field reveals that the strain coupling generated by the 45° twist angle causes different lattice distortion of LSMO, thereby enhancing both the fourfold and uniaxial anisotropy. This work confirms the twisting degrees of freedom for magnetoelectric coupling and opens opportunities for fabricating artificial magnetoelectric heterostructures.Manipulating strain mode and degree that can be applied to epitaxial complex oxide thin films have been a cornerstone of strain engineering. In recent years, lift-off and transfer technology of the epitaxial oxide thin films have been developed that enabled the integration of heterostructures without the limitation of material types and crystal orientations. Moreover, twisted integration would provide a more interesting strategy in artificial magnetoelectric heterostructures. A specific twist angle between the ferroelectric and ferromagnetic oxide layers corresponds to the distinct strain regulation modes in the magnetoelectric coupling process, which could provide some insight in to the physical phenomena. In this work, the LaThe online version contains supplementary material available at 10.1007/s40820-023-01233-z. The ability to control strain in magnetoelectric heterostructures has been one of the key approaches to manipulate a vast range of functional properties, since the strong electronic interactions in complex oxide materials, especially the 3d orbital electrons, make its physical phenomena sensitive to lattice deformations , 2. GeneHowever, the epitaxial growth has fundamental limitations and prevents the unrestricted manipulation of strain in film. Firstly, the maximum strain is determined by lattice mismatch during the epitaxial growth , 9. Seco0.7Sr0.3MnO3  easy axis is observed for 45° Sample, which is significantly different from the uniaxial anisotropy with LSMO [100] easy axis for 0° Sample. Moreover, the in-plane angle-dependent ferromagnetic resonance field (FMR) results exhibit similar magnetic anisotropy, indicating that the 45° rotation of LSMO film on PMN-PT gives rise to different strain modes and lattice distortions from 0° Sample. This study demonstrates the twist angle degree of freedom for performance regulation in magnetoelectric heterostructures and opens up opportunities for fabricating artificial magnetoelectric heterostructures.In this work, an artificial assembly method is used to combine a (001)-oriented ferromagnetic LSMO film and a (011)-oriented ferroelectric single-crystal PMN-PT substrate, which is impossible realized through directly epitaxial growth. LSMO film is epitaxially grown on (001)-oriented SrTiO2 partial pressure was 20 and 30 Pa for the deposition of SAO and LSMO, respectively. Magnetron sputtering technology was used to grow electrodes of 50 nm metal Pt on the upper and lower surfaces of (011)-oriented PMN-PT single-crystal substrates. A resin glue  was used to stick as-grown LSMO films onto the platinized PMN-PT substrates at the twist angle of 45° ± 2° (45° Sample) and 0° ± 2° (0° Sample), respectively. During the transfer process, a continuous pressure was applied to the LSMO/PMN-PT heterostructures, and it was heated at 120 °C for 30 min to ensure complete curing. Finally, the sacrificial layer SAO was dissolved in deionized water for 12 h.PLD was used to epitaxial growth sacrificial layer SAO and ferromagnetic film LSMO in turn on a single-sided polished STO (100) single crystal. The substrates' temperature was 800 °C. The pulsed laser energy was 180 mJ and the frequency was 2 Hz. Meanwhile, the OA high-resolution X-ray diffractogram (XRD) with θ − 2θ scanning  was used to characterize the crystallinity of LSMO/SAO/STO(001) and LSMO(001)/PMN-PT(011). And an atomic force microscope (AFM) (Bruker Dimension Inc.) was used to characterize surface morphology of the LSMO/SAO/STO (001) heterostructure. Meanwhile, the microstructure and epitaxial nature of the film and interface was characterized by scanning transmission electron microscopy (STEM) (JEOL JEM-ARM 200F) with a probe spherical aberration corrector.102 mode with a 360° rotating base. Electric field tuning FMR was explored by in situ poling the PMN-PT (011) along the out-of-plane direction with Pt as electrode.The M–H hysteresis loops were measured by a vibrating sample magnetometer (Lake Shore 7404). An X-band (9.3 GHz) electron paramagnetic resonance system (JES-FA200) was used for FMR measurements. The sample was placed in a rectangular cavity working at the TEl)-oriented diffraction peaks are found by θ–2θ scans, confirming the high crystallinity of LSMO/SAO/STO heterostructures and transferred LSMO films. All the (00l)-oriented diffraction peaks of LSMO films shift to the left, suggesting the increase of the lattice constant c with the disappearance of epitaxial stress after the transfer process. Figure To obtain high-quality LSMO films, SAO (35 nm) and LSMO (70 nm) are sequentially grown on STO 001) substrates using PLD. Then, LSMO/SAO/STO(001) is stuck onto the (011)-oriented platinized PMN-PT. The PMN-PT(011)/LSMO/SAO/STO(001) is immersed in dissolved water to etch the sacrificial SAO layer. Two types of LSMO/PMN-PT are fabricated as shown in Fig.  substratHr) shift indicates the strain relaxation after the transfer, as shown in Fig. Hr decreased by 1028 Oe. Besides, the biaxial in-plane anisotropy and an enhanced Mr/Ms after transfer are observed, as shown in Fig. Hr after strain release, as shown in Fig. The in-plane magnetization curves of the LSMO films before and after the transfer process exhibit the same magnetization features with coercive field only decreasing from 39 to 25 Oe, as shown in Fig. E is applied along the PMN-PT [011], as shown in Fig. Hr–E curve exhibits a loop-like behavior, as shown in Fig. Hr switches at ± 1.6 kV cm−1 and saturates at 7.2 kV cm−1, which provides two nonvolatile states of high and low Hr at 0 kV cm−1.To confirm the magnetoelectric coupling of LSMO/PMN-PT, an electric field Hr shift is synchronously observed in two different heterostructures. The maximum Hr shift along out-of-plane direction is 144.35 Oe for the 0° Sample and 188.48 Oe for the 45° Sample, corresponding to ME coefficient 20.07 and 26.18 Oe cm kV−1. According to the Kittel equations for out-of-plane cases  and LSMO [-110]. This fourfold anisotropy in transferred LSMO film originates from the incomplete release of the biaxial epitaxial tensile strain [Mr/Ms switches to the uniaxial anisotropy with an easy axis toward LSMO [100] for 0° Sample, as shown in Fig. φH = 45°, coinciding with tensile strain and a positive magnetostriction coefficient of LSMO. However, the hard axis is along φH = 160°, which indicates a complex lattice distortion induced by different strain modes.To further explore the in-plane magnetic anisotropy switching, a 7.2 kV cme strain , 36. WheH = 0°) and hard axis (φH = 90°) for 0° Sample. The squareness Mr/Ms dramatically decreases from 0.88 to 0.29 with coercivity synchronously reducing from 72.6 to 25.1 Oe when in-plane magnetic field angular (φH) changes from 0° to 90°. Meanwhile, the coercivity field displays a similar angle-dependent feature with Mr/Ms as shown in . The normalized M–H loop also exhibits a different magnetization feature for 45° Sample. When the magnetic field direction changes from easy axis (φH = 45°) to hard axis (φH = 160°), the Mr/Ms reduces from 0.87 to 0.53 with the coercivity field changing from 40.6 to 36.1 Oe. The differences of the magnetic anisotropy and magnetization feature at the electric field between the two samples further indicate the novel strain-induced magnetoelastic effect in the LSMO/PMN-PT heterostructures.Figure M and external magnetic field H with respect to the sample surface is marked as θ,M φM, and θ,H φH, respectively. The out-of-plane Hr anisotropy for the two samples is not significantly different under + 7.2 kV cm−1, as shown in Fig. Hf, since the fitting values of Hu = 14 Oe and Hf = 33 Oe, which can be attributed to the incomplete release of epitaxial strain and the magnetocrystalline anisotropy of LSMO  and hard axis along LSMO [010], showing a dumbbell-like curve with fitting values Hu = 120 Oe and Hf = 5 Oe, as shown in Fig. Hu = 68 Oe and Hf = 55 Oe imply the coexistence of fourfold and uniaxial anisotropy in 45º Sample. The two different easy axes point to φH = 45° (LSMO [110]) and φH = 140°, while a strong hard axis is along φH = 100°, as shown in Fig. Hu refers to the magnetoelastic energy, which describes the uniaxial stress anisotropy due to piezoelectric strain from the substrate  (φH = 45°). Therefore, the uniaxial magnetic anisotropy rotates 45° and easy axis points to LSMO [110], due to the twist angle. Besides, this twist angle causes a φf = 28° phase shift between uniaxial stress anisotropy and fourfold magnetocrystalline anisotropy, which explains the new type of magnetic anisotropy in 45° Sample. This result further indicates that, by the twisted integration, the specific substrate's piezoelectric strain can produce a twist angle-dependent magnetic anisotropy in the magnetic films. This twist angle degree of freedom could help to discover new physical phenomena in artificial magnetoelectric heterostructures.After applying an electric field of + 7.2 kV cmubstrate , 39. ForIn summary, we applied the epitaxial lift-off technique to stick the epitaxial (001)-oriented LSMO ferromagnetic films onto the (011)-oriented PMN-PT ferroelectric substrates with different twist angles. Due to the 45° twist angle between LSMO and PMN-PT, the biaxial strain from PMN-PT acts on the LSMO layer in different modes, resulting in the coexistence of uniaxial and fourfold in-plane magnetic anisotropy. This twist integration method provides tunability of the twist angle between different layers, allowing a new degree of twist freedom. This study demonstrates the twist angle degree of freedom for performance regulation in magnetoelectric heterostructures and confirms the great research potential of twist angle-adjustable through epitaxial lift-off technology.Supplementary file1 (PDF 501 KB)Below is the link to the electronic supplementary material.


Faujasite-type zeolites are promising adsorbents for CO2 separation due to their high CO2 adsorption capacity. While commonly inert binder materials are used to shape zeolite powders into the desired macroscopic format for application in an adsorption column, here we report the synthesis of Faujasite beads without the use of a binder and their application as CO2-adsorbents. Three types of binderless Faujasite beads (d = 0.4–0.8 mm) were synthesized using an anion-exchange resin hard template. All the prepared beads consisted mostly of small Faujasite crystals, as demonstrated by characterization with XRD and SEM, which are interconnected through a network of meso- and macropores (10–100 nm), yielding a hierarchically porous structure, as shown by N2 physisorption and SEM. The zeolitic beads showed high CO2 adsorption capacity (up to 4.3 mmol g−1 at 1 bar and 3.7 mmol g−1 at 0.4 bar) and CO2/CH4 selectivity . Additionally, the synthesized beads have a stronger interaction with CO2 than the commercial zeolite powder . Therefore, they are also suitable for CO2 adsorption from gas streams in which the CO2 concentration is relatively low, such as flue gas.Biomethane can be isolated from biogas through selective CO Notably, our beads exhibited a steeper adsorption isotherm at low relative pressure compared to the commercial zeolite powder and thus a higher enthalpy of adsorption . This indicates that the interaction of CO2 with our beads is stronger than with the commercial zeolite. This is a particularly useful characteristic for CO2 adsorption from gas streams with relatively low CO2 concentrations, such as flue gas.The binderless beads were compared with extrudates made from commercial zeolite Y. The CO2/CH4 selectivity of the extrudates was superior to that of the beads . Since the extrudates were made from commercial zeolite Y powder, which had relatively large individual crystals (0.35–0.75 mm), it was hypothesized that the diffusion limitations of CH4 within these crystals caused the increased selectivity observed with the extrudates. The beads consist of much smaller zeolitic crystals (30–70 nm) and, therefore, experienced less marked diffusion limitations compared to the extrudates. Additionally, the beads possessed a large amount of meso- and macropores, which is anticipated to be a favourable feature for application in CO2 adsorption because this should lead to fast adsorption/desorption kinetics.The CO2 separation from biogas due to their accessible pore structure, good CO2 adsorption capacity, and good CO2/CH4 selectivity. Additionally, these beads are also attractive for the separation of CO2 from more dilute gas streams due to their relatively high enthalpy of adsorption. To conclude, we synthesized a set of hierarchically porous beads containing Faujasite zeolite domains, which are promising adsorbents for CO


Hantaviridae currently encompasses seven genera and 53 species. Multiple hantaviruses such as Hantaan virus, Seoul virus, Dobrava-Belgrade virus, Puumala virus, Andes virus, and Sin Nombre virus are highly pathogenic to humans. They cause hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome or hantavirus pulmonary syndrome (HCPS/HPS) in many countries. Some hantaviruses infect wild or domestic animals without causing severe symptoms. Rodents, shrews, and bats are reservoirs of various mammalian hantaviruses. Recent years have witnessed significant advancements in the study of hantaviruses including genomics, taxonomy, evolution, replication, transmission, pathogenicity, control, and patient treatment. Additionally, new hantaviruses infecting bats, rodents, shrews, amphibians, and fish have been identified. This review compiles these advancements to aid researchers and the public in better recognizing this zoonotic virus family with global public health significance. Bunyaviridae was officially upgraded to Bunyavirales, and the genus Hantavirus in Bunyaviridae was upgraded to Hantaviridae . Va. Va60]. The attachment and entry of hantaviruses into host cells are mediated by the binding of viral glycoproteins to host cell surface receptors . Some haAfter internalization, virions form clathrin-coated vesicles, which can be formed with clathrin-coated cell membranes . VirionsThe fusion of the viral envelope and endosomal membrane promotes the release of the viral ribonucleoproteins (RNP) complex into the cytoplasm, initiating the transcription and replication of the viral genome . RdRp meN proteins play a variety of roles in the viral replication cycle. They participate in transcription together with viral RdRp to promote mRNA translation . RdRp inThe viral GPC is cleaved in the ER into the Gn and Gc glycoproteins at the conserved pentapeptide motif WAASA , which aThe host mechanisms of innate and acquired immunity in mammals are crucial to inhibiting hantavirus replication .Although hantaviruses can be inactivated by heating at 60 °C for 30 min, organic solvents, hypochlorite solvents, or ultraviolet light , they arUnlike the fact that various bunyaviruses  are transmitted by insects, human hantaviruses are mainly transmitted by rodents ,54,68. HInfected animals can spread hantaviruses to other sensitive animals through aerosols or droplets formed from their excreta or secretions , through the consumption of contaminated food, or through biting and scratching ,70. TheyRodents can also transmit hantaviruses to humans through aerosols or droplets formed from their excreta and through the consumption of contaminated food . Rodent ANDV is currently the only known hantavirus that can be transmitted from human to human. It usually occurs after close contact with an infected person . ANDV maInvestigations have shown that HFRS can be transmitted to animals or humans through the bites of mites ,80. InveHTNV and PUUV antibodies have been detected in cattle, deer, and rabbits. SNV antibodies have been detected in cats and dogs . The rolHuman infections with hantaviruses can cause acute diseases, with the severity varying depending on the viral species and strains ,19,20. H+ T lymphocytes [+ T lymphocytes can be detected in HFRS patients. Elevated IL-6 levels are associated with thrombocytopenia and renal failure [Hantavirus infections in both animals and humans mainly occur in renal or pulmonary endothelial cells (ECs) and macrophages . The maiphocytes . Delayedphocytes . After iphocytes . Activatphocytes . Differe failure . Human leukocyte antigen (HLA) is responsible for presenting viral antigens to T lymphocytes. Genetic susceptibility to severe HFRS disease is related to the HLA type, and different hantaviruses are associated with different HLA haplotypes .Natural reservoir hosts of hantaviruses often exhibit asymptomatic and persistent infection, which may result from the innate immunity of the reservoir hosts . In rodeNlrc3−/− mice infected with HTNV have shown clinical symptoms and pathological changes resembling those seen in patients with HFRS, suggesting a new model for studying the pathogenesis of HTNV. Monkeys can be infected with PUUV through laboratory infection with similar symptoms to NE and can be used as an animal model of NE [The lack of obvious symptoms in natural hosts and the lack of suitable animal models have limited research on the pathogenesis of HTNV and SEOV . Howeverel of NE . Researcel of NE . Epidemiological information and clinical symptoms are useful for the initial diagnosis of hantavirus infections. Confirmation of hantavirus infections should be based on clinical symptoms, epidemiological information, and the detection of the nucleic acids, proteins, and/or antibodies specific to the associated hantavirus . No specific antivirals have been approved for HFRS and HCPS/HPS in the USA or Europe . HoweverProphylactic administration of ribavirin and fapilavir in early infection or post-exposure has shown some efficacy . Both riAntibodies include monoclonal and polyclonal antibodies . MonocloSupportive treatment can be used to maintain fluid and electrolyte balance in HFRS patients, while blood transfusions can control bleeding and improve clotting in DIC cases. For HCPS/HPS patients, oxygen supplementation is usually required, and extracorporeal membrane oxygenation may be employed .The prevention and control of hantaviruses pathogenic to humans rely on education and actions to block the viral transmission and protect humans.Epidemiological surveillance and analysis play key roles in identifying risky regions and factors that contribute to hantavirus transmission. To mitigate this risk, rodenticides, traps, and cats can be employed for the targeted control of rodents. Additionally, cleaning and disinfecting human living environments can prevent the contamination of food and other items with hantaviruses from rodent excreta or secretions. People at high risk of infection with hantaviruses such as those working in agriculture, forestry, or animal husbandry should take protective measures including wearing masks or face coverings . No vaccIn recent years, multiple types of new vaccines for hantaviruses have been investigated in laboratories. For instance, a vaccinia virus recombinant vector vaccine expressing the glycoproteins Gn and Gc of HTNV elicited neutralizing antibodies against both HTNV and SEOV in hamster models, although the vaccine had no cross-protection against PUUV . VariousHantaviridae has been greatly expanded and revised, and research targeting various aspects of this important virus family has made significant progress.From the panorama of hantaviruses depicted above, hantaviruses circulate in various mammals , amphibians, and fish worldwide, and some hantaviruses are highly pathogenic to humans. They usually infect humans through animal-to-human transmission. In recent years, the taxonomy of Hantaviridae can be anticipated. Novel genera or species within this family will be discovered, particularly from novel regions or host species. For instance, the novel hantaviruses Rusne virus from the root vole in Lithuania and the Academ virus from moles in Russia have recently been reported [In the years to come, numerous research advances in reported ,100. Som


Listeria innocua and Escherichia coli was investigated in distilled water (DW) and bacterial culture broth. L. innocua and E. coli at approximately 105 cfu mL−1 were inoculated in PJ samples and incubated at 4, 25, and 37 °C for 0, 6, 24, and 48 h. The bacterial population and pH of culture media were measured at each removal. Results indicated that the antibacterial effect of PJ was dependent upon bacteria species, juice concentration, incubation temperature, and growth medium. Higher juice concentration and incubation temperature resulted in increased antibacterial effects. Bacterial populations were decreased more significantly in DW systems than in the culture broth, while L. innocua was more sensitive to PJ than E. coli in the DW systems. Regardless of PJ concentrations in DW systems, L. innocua, initially inoculated at approximately 105 cfu mL−1, was reduced to undetectable levels at 25 and 37 °C within 24 h. The growth of L. innocua and E. coli was significantly inhibited in bacterial culture broth containing ≥ 20% PJ (p < 0.001). This study provides insight into the potential application of PJ in food and beverage products for food protection.The antibacterial effect of pomegranate juice (PJ) at six concentrations  against  Punica granatum) comprises peel containing polyphenols, minerals, and complex polysaccharides. The edible portion of pomegranates contains organic acids, sugars, vitamins, polysaccharides, polyphenols, and minerals, although the chemical composition of the fruit varies depending on the cultivar, environmental conditions, maturity, and postharvest treatments [The fruit of pomegranate  and bacterial culture broth systems at different temperatures.Adding chemical preservatives to foods can reduce the risk of microbiological contamination. However, consumers have been demanding natural, healthy, and safe foods. The growing demand for developing natural antimicrobials is in response to the perceived risks associated with chemical residues in food ,16,17. Felf life ,18. Ther® Co., Inc., Tokyo, Japan), where approximately 0.3 mL of juice was placed onto the prism surface of the refractometer. Titratable acidity (TA) of PJ was measured with 0.5 N NaOH using the Metrohm Model 848 Titrino Plus  expressed in citric acid equivalents TA (%). The pH of PJ was measured using a pH meter . Total phenolic content  was determined using Folin-Ciocalteu Reagent, and gallic acid was used as standard [Commercially processed 100% PJ (not from concentrate) without any food additives or preservatives was obtained from a local grocery supplier  and stored at 4 °C until use. Soluble solid content of PJ was measured using a digital hand-held refractometer  and E. coli (ATCC10798) were stored at −86 °C in Brain Heart Infusion broth (BHI)  or Tryptone Soya Broth (TSB)  containing 15% glycerol, respectively until use. Prior to the experiment, the working cultures were prepared by sub-culturing 100 µL of L. innocua stock culture in 10 mL BHI and 100 µL E. coli in 10 mL TSB, then incubated at 37 °C for 24 h. The working cultures were centrifuged at 11,500× g for 10 min, supernatants were removed, and bacterial cells were washed with sterile water then centrifuged again. The resulting pellets were made up to a 5 mL volume with sterile water and used for L. innocua or E. coli inoculum. Each inoculum was serially diluted and spiral plated in duplicate on BHI or TSB agar using a WASP 2  spiral plater and incubated at 37 °C for 48 h for enumeration of total population counts of inoculum. For each repeat, the new working cultures were prepared as described above to keep consistent bacterial populations of initial inoculum. v/v). Each tube was then inoculated to obtain an initial inoculum level of approximately 105 cfu mL−1. The samples were shaken well and incubated at 4, 25, or 37 °C to represent cold storage temperature, room temperature, and optimum bacterial growth temperature, respectively. The bacterial population was measured at 0, 6, 24, and 48 h following inoculation. At each sampling time, an aliquot of each sample was serially diluted with 0.1% sterile peptone water , and the appropriate dilution was spiral plated onto the BHI or TSB agar using the 100 µL mode of the spiral plate system, WASP2 for enumeration of L. innocua and E. coli, respectively. After incubation at 37 °C for 24 h, visible colonies of L. innocua and E. coli on the surface of the BHI or TSB agar plates were counted using the aCOLyte colony counter . The antibacterial activity of six concentrations of PJ was investigated in bacterial culture broth and sterilized distilled water (DW) systems. At each concentration level, an appropriate volume of juice was added to the culture broth or DW in a sterile test tube to a final volume of 10 mL. The final concentrations of PJ were 0, 10, 20, 30, 40, and 50% (L. innocua and E. coli), two culture media (bacteria culture broth and DW), six juice concentrations , three incubation temperatures , and four removal times . Bacterial populations in this study were expressed as log cfu mL−1. Data were analyzed using the ANOVA directive and standard errors of mean (SEM) option of GenStat® . All experiments were replicated three times.The experimental design was a completely randomized 2 × 2 × 6 × 3 × 4 factorial with two bacteria species , titratable acidity (TA) was 0.91% ± 0.10, pH value was 3.32 ± 0.07, and total phenolic content  was 1717.04 mg GAE lv/v) in the BHI broth had a significant inhibitory effect on the growth of L. innocua  at 6 and 24 h incubation b. Again,L. innocua in 0, 10, and 20% PJ over time  within 24 h at PJ concentrations ≥ 10% in distilled water  over time. The 50% PJ concentration resulted in the greatest reduction of E. coli to an undetectable level after 48 h incubation.At 37 °C incubation temperature, the concentrations of PJ at 20, 30, 40, and 50% in the TSB suspension had a significant inhibitory effect on the growth of E. coli a. E. colE. coli population was similar to that of 37 °C for the concentrations of 0 and 10%  over 48 h (E. coli population significantly decreased over time compared with the control (p < 0.001). Furthermore, E. coli populations decreased to undetectable levels (<10 cfu mL−1) after being treated with PJ at concentrations ≥ 20% for 48 h.The population of ver 48 h c. HoweveE. coli inoculated in the control or 10% juice suspension over 48 h in distilled water  at 4, 25, and 37 °C  were investigated, which were different from the research reported previously. PJ showed antibacterial effects against Gram-positive L. innocua and Gram-negative E. coli, while both bacteria were inhibited and killed more effectively in DW systems than in culture broth systems. PJ has intrinsic characteristics with low pH values and high titratable acidity that create a hostile environment for bacterial growth and survival. Microbial response to acidic pH is a major factor controlling the growth and survival of bacteria in foods and acidic body environments [L. innocua was found to be more sensitive to PJ than E. coli, as significant L. innocua reductions (p < 0.001) were shown following the additions of PJ for 6 h at three temperatures tested in DW systems. Gahan et al. [L. monocytogenes is at pH 3.5 without the treatment of adaptive acid tolerance response by using nutrient broth systems. The L. innocua did not tolerate the acidified conditions with limited nutrients. E. coli could better tolerate the lower pH in comparison to L. innocua. Miller and Kaspar [E. coli O157:H7 could be detected in apple cider with a pH of 3.6 stored at 8 °C and was shown not to be destroyed within the shelf life of apple cider at pH 2 to 4. Nikaido [Listeria spp. and E. coli may lead to different effects when the bacteria are subjected to antimicrobial compounds [Pomegranate fruits are known as a good source of healthy antioxidants, including polyphenols and anthocyanin ,20. Whilronments . L. innon et al.  reported Nikaido  reported Nikaido , the celompounds . −1 in this study), also contribute to the antibacterial actions. The primary polyphenols of 4-Hydroxy-3-Methoxybenzoate, epicatechin, catechin, rutin, ferulic acid, P-coumaric acid, and cinnamic acid in pomegranate juice concentrate (PJC) were determined [Streptococcus mutans and Aeromonas hydrophila. Wu et al. [Salmonella enterica, indicating that the level of Salmonella population reduction correlated positively with the total polyphenolic content and titratable acidity in the treatment system. Phenolics with hydrophobicity could act efficiently at the bacterial membrane–water interface by embedding in the membrane, thereby impairing the cell membrane [+-ATPase required for ATP synthesis. This damage may make the bacteria more susceptible to the phenolic compounds in the PJ. Türkyilmaz et al. [E. coli, P. aeruginosa, and C. neoformans with IC50 values lower than 15 µg mL−1. It is worth noting that the higher concentration of PJ showed greater antibacterial effects than the lower concentration of PJ at the same pH level . Bacteritermined . PJC demu et al.  studied membrane . Apostolmembrane  and Lin membrane  reportedz et al.  reportedz et al.  determinL. innocua and E. coli. Higher incubation temperatures resulted in stronger bactericidal effects. In this study, both bacteria were less affected by PJ at 4 °C. With abundant nutrients and optimum temperature, the newly formed cells of the bacteria are active and multiply quickly. The cell wall of newly formed cells is more easily destroyed than that of older cells [Different treatment temperatures had impacts on the antimicrobial activities of PJ against er cells . Aronssoer cells  reporteder cells .L. innocua and E. coli in bacterial culture broth systems, suggesting that PJ has antimicrobial potential in the food system. Xylia et al. [E. coli and L. monocytogenes population on shredded carrots, while total carotenoids, ascorbic acid, total soluble solids, and titratable acidity were not different after six days of storage of samples. Pomegranate has been used in the food industry, including juice for drinking and its extract for extending the shelf life of foods. PJ and its extracts present high antioxidant and antimicrobial activity, which make them excellent candidates as natural additives or active compounds in the food and food packaging industries [Our study demonstrated the antimicrobial effects of PJ against a et al.  also repdustries . To bettL. innocua and E. coli were more effectively killed in the DW systems than in their culture broth, while L. innocua was more sensitive to PJ compared to E. coli. Low pH value and chemical compounds in PJ contributed to the antimicrobial effect. This study provides useful information for the food industry to develop pomegranate juice and/or its extracts as healthy food/beverage as well as natural food additives for food protection. The antimicrobial effects of PJ were dependent upon the juice concentration, culture conditions , as well as bacterial species. Increased juice concentration and treatment temperature resulted in a greater antibacterial effect.


Monkeypox (Mpox) is a contagious illness that is caused by the monkeypox virus, which is part of the same family of viruses as variola, vaccinia, and cowpox. It was first detected in the Democratic Republic of the Congo in 1970 and has since caused sporadic cases and outbreaks in a few countries in West and Central Africa. In July 2022, the World Health Organization (WHO) declared a public-health emergency of international concern due to the unprecedented global spread of the disease. Despite breakthroughs in medical treatments, vaccines, and diagnostics, diseases like monkeypox still cause death and suffering around the world and have a heavy economic impact. The 85,189 reported cases of Mpox as of 29 January 2023 have raised alarm bells. Vaccines for the vaccinia virus can protect against monkeypox, but these immunizations were stopped after smallpox was eradicated. There are, however, treatments available once the illness has taken hold. During the 2022 outbreak, most cases occurred among men who had sex with men, and there was a range of 7–10 days between exposure and the onset of symptoms. Three vaccines are currently used against the Monkeypox virus. Two of these vaccines were initially developed for smallpox, and the third is specifically designed for biological-terrorism protection. The first vaccine is an attenuated, nonreplicating smallpox vaccine that can also be used for immunocompromised individuals, marketed under different names in different regions. The second vaccine, ACAM2000, is a recombinant second-generation vaccine initially developed for smallpox. It is recommended for use in preventing monkeypox infection but is not recommended for individuals with certain health conditions or during pregnancy. The third vaccine, LC16m8, is a licensed attenuated smallpox vaccine designed to lack the B5R envelope-protein gene to reduce neurotoxicity. It generates neutralizing antibodies to multiple poxviruses and broad T-cell responses. The immune response takes 14 days after the second dose of the first two vaccines and 4 weeks after the ACAM2000 dose for maximal immunity development. The efficacy of these vaccines in the current outbreak of monkeypox is uncertain. Adverse events have been reported, and a next generation of safer and specific vaccines is needed. Although some experts claim that developing vaccines with a large spectrum of specificity can be advantageous, epitope-focused immunogens are often more effective in enhancing neutralization. In recent years, there has been cause for concern due to a rapid and unprecedented pandemic of Mpox infections in several countries all over the world. Rodents and primates are the hosts for Mpox, a zoonosis  with symptoms like smallpox but less severe. When the virus was initially identified in monkeys in a Danish laboratory in 1958, the term “monkeypox” was coined ,3. ThereThe current research implies that monkeypox may spread in three different ways: from person to person, via direct contact with infected organisms, and from animals to people. It is well established that animals can pass the Mpox virus on to humans . The majThe Mpox virus is categorized as a category of the genus Orthopoxvirus under the family Poxviridae . Macropi. It was believed that native prairie dogs housed alongside rats from Ghana introduced in Western Africa were the main source of the pandemic. This is because most infected individuals became ill after coming into contact with pet prairie dogs [n = 295 cases) compared to week 2 (9 January–15 January) (n = 302 cases). The bulk of cases recorded in the last four weeks were from the Americas region (77.7%) and the African region (13.9%). The United States of America , Brazil , Spain (n = 7518), France (n = 4114), Colombia (n = 4066), the United Kingdom (n = 3735), Peru (n = 3723), Mexico (n = 3696), Germany (n = 3690), and Canada (n = 1460) are the ten most afflicted nations worldwide listed in The Mpox virus was first reported in 1958 in laboratory monkeys employed for research purposes at State Serum Institutes in Copenhagen, Denmark, as well as in Africa ,43. Humarie dogs  In the srie dogs . Since 2rie dogs . In 2018rie dogs . Over thrie dogs ,49. Mpoxrie dogs ,51,52. Orie dogs . By 29 Jrie dogs . Accordirie dogs . This scrie dogs ,57. GlobMpox is a virus belonging to the Orthopoxvirus genus and has a clinical presentation similar to smallpox. Its incubation period in humans typically ranges from four to 14 days but can be as long as 21 days. The disease begins with a febrile prodrome, which is accompanied by headache, muscle aches, backache, exhaustion, sweats, and fatigue. One to three days after the onset of fever, a rash appears on the face, inside the mouth, and on the hands, feet, chest, genitals, anus, and eyes. This rash begins as a flat macula and then becomes a papule before forming a vesicle filled with clear liquid. This clear liquid then turns into a yellowish liquid and forms pustules. Once the pustules, crusts, and lesions fall off, the patient is no longer considered infectious. However, scarring from the rash is a common outcome of an infection. More severe complications such as pulmonary distress, bronchopneumonia, ocular infections, corneal scarring, and even permanent damage, as well as lymphadenopathy, may also occur. It can be difficult for clinicians to differentiate Mpox from other viral or nonviral diseases due to its nonspecific clinical presentations. Therefore, laboratory diagnosis is imperative . In ordePhylogenomic studies indicate that there are at least three distinct clades of the Mpox virus—Clade 1, associated with West Africa; Clade 2, connected to the Congo Basin; and a novel Clade 3, proposed following the 2022 European outbreak. Although the clinical appearance of smallpox and Mpox viruses are similar, the three clades suggest a distinct evolutionary divergence . The MpoIn order to prevent the spread of the Mpox virus and protect people from it, there are multiple steps that must be taken. Vaccines are the most effective way to achieve this goal, yet unfortunately, there is currently no specific vaccine for the Mpox virus. However, research has revealed that the same smallpox vaccine that was used to protect people against smallpox may be effective in protecting against the Mpox virus as well . PreviouA total of 85,189 cases of Mpox infection as of 29 January 2023 have alarmed the world. Historically, immunization against the vaccinia virus could safeguard against Mpox; however, since smallpox was eradicated, this type of vaccination has ceased. Consequently, therapeutic options for those already infected are of considerable importance . No antiCurrently, operational research faces challenges in understanding the dynamics of monkeypox transmission and control due to limited resources for detailed case investigations and contact follow-up in affected communities. A serious issue is the lack of adequate diagnostic facilities in laboratories. The difficulty in diagnosing the Mpox virus arises from the insufficiency of laboratory-diagnosis capacity and access, making it challenging to identify any underlying etiology. To comprehend the epidemiology and subclinical infection among contacts in communities, a seroprevalence study is crucial. However, currently available serological assays are generic orthopox tests, and they cannot specifically identify the Mpox virus due to cross-reactivity between the Mpox and smallpox viruses. Hence, it is challenging to distinguish between Mpox-virus infection and prior smallpox vaccinations or other orthopoxvirus infections. Moreover, these assays are not available on the marketplace. Data collected from Nigeria reveal that approximately 20% of 70 monkeypox-negative patients with a rash illness that had similar antigens also had orthopox antibodies. In addition, there is no specific antiviral treatment for Mpox. Treatment is primarily supportive, focusing on managing symptoms such as fever and rash. Vaccination with the smallpox vaccine can provide some protection against Mpox, but the vaccine is not widely available in many countries. Further research, including molecular and genomic approaches, is necessary to identify other orthopoxviruses transmitted in human and animal populations.Mpox is a viral disease that is closely related to smallpox and primarily found in remote parts of Central and West Africa. There is currently no specific vaccine for Mpox, but the smallpox vaccine provides some protection against the disease. However, the smallpox vaccine is no longer routinely administered, and many younger people in Africa may not have received it. Several vaccines are being studied for their potential effectiveness against Mpox, including live-attenuated Mpox vaccines, DNA vaccines, and recombinant vaccines. Synthetic peptide-based prototype vaccines have also shown promise in preclinical studies, and researchers are investigating the use of mRNA vaccines for booster purposes in those who have received the mRNA vaccine for COVID-19. Overall, the development of effective vaccines for Mpox is an ongoing area of research, and scientists are working to develop new vaccines that can protect against this rare but potentially serious disease.


In regard to the analysis pertaining to the dynamic fracture mechanism of marble under the influence of the freeze-thaw environment that characterizes the plateau, the aforementioned experimental results exhibit considerable significance.To study the effects that the perennial freeze–thaw environment exerts on the dynamic mechanical properties of marble, which characterizes the Qinghai-Tibet Plateau, impact tests were conducted, and saturated marble was utilized; thus, we analyzed the effect of different loading rates on its dynamic compressive strength, fragmentation pattern, and energy-absorbing density. The results indicate the following: (1) When 42.02s Due to years of freeze–thaw cycles, the dynamic mechanical characteristics of the rocks that characterize alpine environments have changed considerably. Furthermore, throughout the mining process, groundwater is frequently encountered, and an immense amount of rock becomes saturated with water. Because they are a non-homogeneous material, rocks exhibit numerous small fractures, which permeate their internal structure. Therefore, it is crucial to analyze the dynamic mechanical properties of the rocks that characterize a saturated state, which pervade mining plateaus, earthquake prediction, tunnel and rock explosion –5.Numerous local and international scholars have immensely analyzed saturated rocks; thus, the physical and mechanical properties of saturated rocks have been somewhat revealed. A.M. Rubin  analyzedTo reveal the microscopic mechanisms and critical phenomena that characterize the softening of saturated soft rocks, Cuiying Zhou , 15 utilThe aforementioned study indicates that with respect to rocks, water affects the physical and mechanical properties, energy dissipation properties, and fractal fracture. Herein, uniaxial impact tests, which utilize different loading rates, were conducted on dry and saturated marble; the marble was obtained from the Niu Kutou mining area that is situated in the Qinghai plateau region, and using a split Hopkinson com-pression bar, the energy transfer law of marble under different strain rates was explored, the fractal law of marble mass after impact crushing was analyzed, the relationship between the energy dissipation that characterizes marble crushing under impact load-ing and fractal dimension was constructed, and the dynamic strength characteristics of marble in different regions was compared; thus, a theoretical guidance for comprehending the dynamic mechanical properties of saturated rocks, which characterize the plateau freeze-thaw environment, was developed.In regard to this test, the marble that was utilized was obtained from the Niu Kutou mining area; the mining area is located on the north slope of Qimantag Mountain, which is located in the northeastern part of the Qinghai-Tibet Plateau, a typical alpine area with an average annual temperature of 4.3°C. The groundwater veins are somewhat developed, and they belong to the saturated section. To ensure that the non-parallelism and non-perpendicularity of the specimen ends were less than 0.02 mm, the specimen ends were sanded and polished , 24.The basic mechanical parameters of the specimen are depicted in In regard to this test, the kinetic SHPB test system that was recommended by the International Society of Rock Mechanics was utilized, and in regard to the determination of rock kinetic parameters, this equipment has been widely utilized , 23. FigThe initial air pressure was set at 0.25 MPa, and by gradually increasing the gradient by 0.05 MPa, the impact test was performed on saturated marble and dry marble.VT) with the incident voltage and reflected voltage (VI+VR) in the original waveform, dis-carding the unqualified data, and making up the corresponding test, this study examines the stress wave balance. VI+VR waveform approximates the VT waveform, which indicate the test voltage balance.Based on the one-dimensional stress propagation theory, we can determine the rationality of the SHPB test, and to ensure the reliability of the test, the stress balance principle must be followed , 26. TheThree sets of impact tests were conducted at each air pressure, and for analysis, the data that was obtained from the three tests was averaged. Using the three-wave method, the stress–strain rates were calculated –29. UsinThe stress–strain curves pertaining to the uniaxial impact tests that were conducted on dry marble and saturated marble are depicted in As depicted in It is indicated that the dynamic compressive strength of the rock specimens that are subjected to both states exhibits a satisfactory linear relationship with the average strain rate. After water saturation, the strength of the rocks weakened significantly, and after water saturation treatment, the dynamic compressive strength of the marble was reduced by approximately 10.42%~22.61%, which may be occasioned by the following: after the water-filled test, the water reduces the cementation between the mineral particles that constitute the rock, and when the sample is subjected to external loading, the free water that is contained inside the specimen travels to the crack tip and forms a “water wedge”, which accelerates the crack expansion and damages the specimen. Several of these factors may have contributed to the following observation: the dynamic compressive strength of saturated marble is considerably lower than that of the dry state.DIF), the ratio of dynamic peak stress to static peak stress, is introduced to quantify the rate dependence of marble strength -lgr-fitting curve, set the slope as b, where b = 3-D; D denotes the fractal dimension; r denotes the characteristic value of the particle size; and rz denotes the maximum diameter of the fragment. Take the double logarithm on both sides of the above formula to get the following formula.The grading standard sieve is selected, and the pieces are classified into ten kinds of particle-size grades, namely 0~0.3mm, 0.3~0.5mm, 0.5~1.0mm, 1.0~2.5mm, 2.5~5.0mm, 5~10mm, 10~15mm, 15~20mm, 20~25mm, and >25mm. Weigh the pieces that exhibit different particle sizes after screening, the percentage of each particle size is calculated; furthermore, by plotting the distribution curve of rock crushing bulk at different strain rates, the fractal dimension is calculated, as expressed in the following equation :M(r)M(M(r)/M(z)]-lgr curves pertaining to the dry and saturated state marble are depicted in The fragmentation bulk distribution lg[To study the fractalization and energy dissipation laws of marble under impact loading, draw a scatter diagram of strain rate and fractal dimension and perform fitting analysis on it. The results are depicted in -1, the fractal dimension of the dry state marble is greater than that of the saturated marble; however, the growth rate of the dry state marble is lower than that of the saturated marble, which indicates that during this stage, the inhibitory effect that water exerts on marble fragmentation is weakened; When 42.02s-1 ≤ -1, the fractal dimension of satiated marble and that of dry marble approximate each other, which indicates that in regard to the aforementioned strain rate range, water exerts a low influence on the fractal fractality of marble; When -1, the fractal dimension of dry marble is larger than that of saturated marble, and the difference between the two states becomes larger, which indicates that during this stage, the effect of water on the fragmentation degree of marble is highly significant.Macroscopically, the degree of fragmentation pertaining to the saturated marble is lower than that of dry marble; under the action of water, the absorbed energy that is transformed by the energy that is transmitted by the stress wave into the interior of the specimen is preferentially distributed to the main cracks inside the rock specimen; the expansion of fine cracks is inhibited, and nascent cracks cannot be effectively produced. The rock becomes damaged, which leads to the dissipation of energy, and this energy fails to comprehensively act on the crack expansion; thus, more large pieces are formed, which leads to the reduction of the fractal dimension value.To reveal the correlation between the fractal dimension and dynamic compressive strength and the energy-absorbing density pertaining to different states of marble damage, the fractal dimension, compressive strength and energy absorption density of marble are fitted with a quadratic function with good effect and the results are depicted in Based on the results of dynamic impact test, which considered the marble that is subjected to dry conditions and the marble that is subjected to saturated conditions, the dynamic parameters and damage fractal characteristics of marble are obtained. The relationship between the two specimens is analyzed, and the following conclusions are obtained:Based on the aforementioned comparative analysis, the Niu Kutou mining area that is located in Qinghai is situated in the highland alpine region, and the marble that characterizes the mining area exhibits different impact dynamics, which distinguishes it from other non-alpine marbles under the effect of multiple freeze–thaws; furthermore, the freeze-thaw effect exerts an apparent weakening effect on the marble.The mechanical properties of saturated marble are the result of the interaction of in-ternal free water weakening and strengthening. When the strain rate is low, the weakening effect of water on rock strength is still dominant, showing that the strength of saturated marble is much lower than that of dry marble. As the strain rate increases, the weakening effect of water gradually decreases. In theory, when the strain rate reaches a threshold, the weakening effect of water on the rock can be completely canceled out, and the saturated rock starts to be stronger than the dry rock.-1 or -1, the degree of fragmentation of dry marble is greater than that of the saturated marble, and the fractal dimension is larger; And when 42.02s-1 ≤ -1, the fractal dimension of the saturated marble is larger than that of the dry marble, which indicates that water exerts different effects on the degree of impact crushing that is exhibited by marble in different strain rate ranges.The impact crushing bulkiness of dry marble and saturated marble exhibits similar fractal characteristics, and there is a considerable positive correlation between D and With respect to both the dry and saturated states, there is a positive correlation between the energy-absorbing density of marble and its fractal dimension, and the energy-absorbing density of the saturated marble is greater than that of the dry marble under a constant fractal dimension. In addition, in regard to both states, when the fractal dimension increases, the energy-absorbing densities of the marble converge, which indicates that when the fractal dimension increases, the influence of water on the energy-absorbing properties of marble decreases.


Cowpea chlorotic mottle virus (CCMV) and brome mosaic virus (BMV) are naked plant viruses with similar characteristics; both form a T = 3 icosahedral protein capsid and are members of the bromoviridae family. It is well known that these viruses completely disassemble and liberate their genome at a pH around 7.2 and 1 M ionic strength. However, the 1 M ionic strength condition is not present inside cells, so an important question is how these viruses deliver their genome inside cells for their viral replication. There are some studies reporting the swelling of the CCMV virus using different techniques. For example, it is reported that at a pH~7.2 and low ionic strength, the swelling observed is about 10% of the initial diameter of the virus. Furthermore, different regions within the cell are known to have different pH levels and ionic strengths. In this work, we performed several experiments at low ionic strengths of 0.1, 0.2, and 0.3 and systematically increased the pH in 0.2 increments from 4.6 to 7.4. To determine the change in virus size at the different pHs and ionic strengths, we first used dynamic light scattering (DLS). Most of the experiments agree with a 10% capsid swelling under the conditions reported in previous works, but surprisingly, we found that at some particular conditions, the virus capsid swelling could be as big as 20 to 35% of the original size. These measurements were corroborated by atomic force microscopy (AFM) and transmission electron microscopy (TEM) around the conditions where the big swelling was determined by DLS. Therefore, this big swelling could be an easier mechanism that viruses use inside the cell to deliver their genome to the cell machinery for viral replication. Cowpea chlorotic mottle virus (CCMV) and brome mosaic virus (BMV) are naked plant viruses, and both are members of the Bromoviridae family. CCMV is a virus that commonly infects Vigna unguiculata [Bromus inermis and other grasses and can be found almost anywhere where wheat is grown. It is also one of the few grass viruses that infects dicotyledonous plants, such as soybean [monocotyledonous plants, such as barley and other plants of the Gramineae family. The CCMV and BMV viruses are composed of an icosahedral protein capsid that is approximately 28 nm in diameter [uiculata ,2, while soybean ; howeverdiameter . Their cdiameter . The RNAdiameter . On the diameter . RNA-2  in the presence of Mgutrality ,17. At vutrality ,19,20.The proteins of most viruses assemble in an icosahedral or helical symmetry around their genome. This is a self-assembly process that has also been observed only with the protein subunits without their genome , and theLlauro et al.  conducteUnderstanding the virus capsid swelling is essential, especially for RNA viruses, because it can help us comprehend how a virus disassembles and/or delivers its genetic material within the cells, as this is a crucial step in the virus replication process. Understanding how this process occurs will give us more information about the function of the capsid within the host cell and how it helps in the release of the genetic material protected by the capsid . In thisVigna unguiculata) plants and purified by ultracentrifugation, according to Lavelle et al. [Hordeum vulgare) and was purified by ultracentrifugation following the method of Michel et al. [The CCMV was obtained from infected California black eye pea  or quasi elastic light scattering (QELS); it is a common, non-invasive technique for measuring particle size and size distribution, typically in colloidal and nano colloidal suspensions. Virus sizes were measured with a Zetasizer NanoZS . Samples were prepared by taking aliquots of the purified virus and then equilibrating them by dialysis against a buffer at a given pH and ionic strength for 24 h, using an 8–12 kDa membrane. Virus samples were prepared in a pH range from 4.8 to 7.6, increasing the pH by 0.2 from sample to sample using the appropriate buffers. For a low pH (in the range from 4.8 to 5.8), we used citrate buffers and phosphate buffers for a higher pH (from 6.0 to 7.6) . The virAtomic force microscopy (AFM) images were acquired with a MultiMode V8 SPM NanoScope microscope  in a liquid cell at room temperature. The pH and ionic strength conditions for which the experiments were performed were determined from DLS measurements. The images were obtained using the ScanAsyst fluid mode with 512 pixels/line settings and a scanning rate of 0.5 Hz. To ensure the adherence of the virus to the mica surface in solution, it was treated by two methods to change the mica surface charge from a negative to a positive charge, so the negatively charged capsid would adhere to the mica surface. In the case of CCMV, a freshly cleaved mica surface was treated with 10 μL of poly-L-lysine solution  to alter its surface charge, serving as the substrate . FollowiIn the case of the BMV virus particles, a freshly cleaved mica was first suspended in a magnesium acetate solution (33 mM) overnight to change the surface charge from negative to positive, and after this, it was placed on the AFM piezo scanner, and the liquid cell was mounted afterward. The experiments were carried out in the liquid cell, which was filled with the BMV virus solution in phosphate solution at pH 7 and with an ionic strength of 0.1. Subsequently, it was allowed to stand for two hours to allow the BMV virus particles to adhere to the mica surface through electrostatic interactions. Images were also obtained using the ScanAsyst fluid mode with a setting of 512 pixels/line and a scanning rate of 0.5 Hz. The force applied to both samples, CCMV and BMV, was set to the lowest possible value to avoid surface damage. The cantilever had a nominal tip curvature radius of 2 nm, a spring constant of approximately 0.7 N/m, and a nominal resonance frequency of 150 kHz. The AFM analysis was carried out with the measurement of the height of 100 particles; this study was carried out in the equipment system.The size and morphology of the virus were also characterized by TEM using a JEM-200CX  transmission electron microscope (TEM) at 100 kV equipped with a digital camera . As mentioned before, the samples were prepared under the conditions where big swelling was detected by the DLS experiments, and then the grid was stored in a desiccator overnight. A 6 µL drop was deposited on a copper grid (300 mesh) that was previously coated with parlodion and carbon. After 1 min, the excess sample was removed with a filter paper, and then the sample was negatively stained with uranyl acetate for samples at pH < 6, and for pH > 6, with a phosphotungstic acid solution. The tungstic acid was prepared at the same pH of the sample; this is important because it can be used in samples with a pH greater than 6 . The imaAs mentioned above, the swelling process of a virus is something that has been proposed as a possible mechanism by which the virus delivers its genetic material to the cell translation machinery. To study this phenomenon, a different series of microdialysis was carried out on two different viruses from the same family under different ionic strength and pH conditions, finding quite interesting results.To evaluate the size for CCMV and BMV, we first used DLS over a pH range of 4.8 to 7.6 and ionic strengths (I) of 0.1, 0.2, and 0.3, and all the virus samples had a concentration of 0.3 µg/µL. It is important to mention that the DLS shows the hydrodynamic diameter of the capsid, which makes the measurement ~2 nm higher than that observed by TEM. This is due to the fact that water molecules are dipolar, which favors their association with negatively charged surfaces, in this case, the capsid surface. The presence of water molecules increases the size of the capsid measured by DLS, while the measurement in TEM is performed in a vacuum.Surprisingly, in The evaluation of the swelling of the BMV was carried out in a similar way as the CCMV virion by DLS. The results obtained showed that for an ionic strength of 0.1 and 0.2, the greatest swelling of the virion occurred at a pH of 7.6 and 6.8, respectively, where the swelling was about 25%. For the ionic strength of 0.3, bigger swelling was observed at a pH of 7.0 and 7.6, where the swelling was about 20%. As well as in the case of CCMV, in most cases of pH and ionic strength, swelling was about 10% of the reference value of the virion hydrodynamic diameter.To verify these results, samples were prepared to be observed by TEM under the conditions in which large swelling was detected by DLS and compared to the normal size of the virion, although the TEM results also support that this swelling does not occur in a uniform way. Instead, we see a deformation of the virus capsid, as shown in Furthermore, we characterized the swelling of CCMV and BMV virions by AFM in a liquid cell. The experiments in These results were compared with those of Wilts et al. , where tIt was possible to verify that the CCMV and the BMV present a change in their capsid size when exposed to different conditions of I and pH, resulting in the swelling of the capsid. In addition, we have found that there are specific pH levels and ionic strength conditions under which much greater swelling occurs than reported before. The structural homology of BMV and CCMV capsids is almost identical, but there is a significant difference in the amino acid sequences. Approximately 70% of the sequence is conserved at the protein level. While the beta barrel jelly roll fold is present in many capsids, the bromovirus capsid structure appears to have evolved from other types of capsids based on structural homology. It is possible that BMV and CCMV capsid proteins have a higher structural similarity to kexin-like proteases, unlike other capsid proteins found in nature, and specifically the P domain of these proteases . TherefoIn fact, a key question is whether, given their reversibility, the capsids remain intact with their RNA intact or whether, upon reversion, the virus undergoes changes or loses the RNA. We attempted to address this question using the retardation gel in We assume that in the Wilts et al.  experimeMost vectors transmitting plant viruses are insects, which spread the virus to a new host plant after eating infected leaves and moving to another uninfected plant and eating new leaves . TypicalHowever, we observed a swelling increase much greater than 10 percent for both CCMV and BMV, although BMV appears to swell significantly over a broader pH range than CCMV. It is possible that for BMV, the change is more significant due to the fact that the capsid is less rigid in terms of its structure, more malleable in terms of its form, and more susceptible to changes, in accordance with the experimental studies carried out on BMV . A simplFrom these studies, it has been suggested that ribosomal factors may only require the 5′ end of the RNA, such that the RNA can be extracted from the capsid, which is commonly referred to as co-translational disassembly. The capsid is not required to disassemble immediately upon translation initiation, as has been observed in vitro . PrefereWe show that the CCMV and BMV present high magnitudes of swelling at different ionic strengths and pHs, but more importantly, we show that swelling can be much greater than 10%, as previously reported. It might be that a swelling of around 10% might not be enough for RNA delivery to the cell translational machinery. Whereas a swelling of 20 to 35%, as reported here, could make the viral RNA more accessible for translation. Our work could help better understand how viruses, once inside the host cell, deliver their genetic material to the cellular machinery due to a greater capsid swelling than previously thought, although the detailed mechanisms of this process need more studies.