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Search Results (361)

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10 pages, 889 KiB  
Review
Mpox and Surgery: Protocols, Precautions, and Recommendations
by Nikolaos Kamaratos-Sevdalis, Islam Kourampi, Nazli Begum Ozturk, Anna C. Mavromanoli and Christos Tsagkaris
Microorganisms 2024, 12(9), 1900; https://doi.org/10.3390/microorganisms12091900 - 15 Sep 2024
Viewed by 270
Abstract
Mpox, also known as Monkeypox, is an infectious disease known to spread via direct contact and fomites, which poses a significant contagion risk in surgical settings and may increase the challenges already posed by COVID-19. Within the three years following the outbreak of [...] Read more.
Mpox, also known as Monkeypox, is an infectious disease known to spread via direct contact and fomites, which poses a significant contagion risk in surgical settings and may increase the challenges already posed by COVID-19. Within the three years following the outbreak of Mpox, we conducted a review of the impact of Mpox on surgical practice. We searched Pubmed/Medline and Scopus, focusing on original studies and case reports in English or German. Our search terms included “Mpox”, “Monkeypox”, and “Surgery”. Out of 60 clinical or epidemiological studies, as well as expert opinions, brief reports, and pertinent literature reviews, eight were included after full-text assessment. We also incorporated two pertinent literature reviews, including a total of 10 papers, in this analysis. The main topics addressed by the literature are 1. manifestations of Mpox for surgical consideration or urgent management, for which it is important to consider whether a surgical approach is needed to address long-term Mpox-related lesions and 2. infection control in surgical settings, especially considering its impact on elective surgery and the well-being of healthcare workers. Mpox could affect surgical services and access to operating theaters. Unlike COVID-19, Mpox, compared to initial concerns, has not substantially compromised surgical delivery. However, limited reports exist on the surgical impact of Mpox. It is crucial to involve surgeons in Mpox diagnosis, educate surgical practitioners on its mimicry of common surgical conditions, enhance infection control during surgery, and ensure access to corrective surgery as a means of tackling the stigmatization associated with Mpox and sexually transmitted diseases in general. Full article
(This article belongs to the Special Issue Monkeypox—Current Knowledge and Future Perspectives)
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<p>Mpox and surgery: protocols, precautions, and recommendations. Literature search and selection flowchart.</p>
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<p>Countries of origin of the included studies.</p>
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11 pages, 1647 KiB  
Article
Update of the Genetic Variability of Monkeypox Virus Clade IIb Lineage B.1
by Fabio Scarpa, Ilenia Azzena, Alessandra Ciccozzi, Francesco Branda, Chiara Locci, Maria Perra, Noemi Pascale, Chiara Romano, Giancarlo Ceccarelli, Giuseppe Terrazzano, Pier Luigi Fiori, Massimo Ciccozzi, Marco Casu and Daria Sanna
Microorganisms 2024, 12(9), 1874; https://doi.org/10.3390/microorganisms12091874 - 11 Sep 2024
Viewed by 917
Abstract
From 1 January 2022 to 31 May 2024, the World Health Organization (WHO) reported 97,745 laboratory-confirmed Mpox cases, including 203 deaths, across 116 countries. Despite a 2.3% decrease in new cases in May 2024 compared to April 2024, significant regional variations persist. The [...] Read more.
From 1 January 2022 to 31 May 2024, the World Health Organization (WHO) reported 97,745 laboratory-confirmed Mpox cases, including 203 deaths, across 116 countries. Despite a 2.3% decrease in new cases in May 2024 compared to April 2024, significant regional variations persist. The African Region reported the highest proportion of new cases, while other regions experienced mixed trends. Phylogenomic analyses of the Mpox virus Clade IIb lineage B.1 reveal stable genetic variability with minimal diversification. The Bayesian Skyline Plot indicates a generally stable viral population size with a modest peak in late 2023, followed by a decline. In general, the data indicate that the MPXV outbreak is primarily localized within a few consistent geographic clusters. The virus’s evolution is relatively slow, as indicated by its stable genetic variability, and Clade IIb lineage B.1 does not currently show signs of rapid genetic changes or population growth. The current low level of genetic diversity should not lead to complacency. Ongoing genomic surveillance is essential for effective outbreak management and understanding. This monitoring is crucial for identifying any shifts in the virus’s behavior or transmission, allowing for prompt public health responses and adjustments. In addition, continued vigilance is necessary to detect any new variants that might influence the outbreak’s trajectory. Full article
(This article belongs to the Special Issue Monkeypox—Current Knowledge and Future Perspectives)
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<p>Highlights of all clades and lineages of MPXV collected between June 22 and July 2024 in the time-scaled phylogenetic tree constructed with 3822 complete genomes (last updated 31 July 2024. (<b>A</b>) Terminals labeled according to the GISAID Clade of belonging. (<b>B</b>) Terminals labeled according to the country of origin. The figure has been edited using the software GIMP 2.8 (available at <a href="https://www.gimp.org/downloads/oldstable/" target="_blank">https://www.gimp.org/downloads/oldstable/</a>, accessed on 7 August 2024).</p>
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<p>(<b>A</b>) Bayesian Skyline Plot (BSP) for the MPXV Clade IIb lineages B.1. This plot shows the genetic variability and thus the effective viral population size (<span class="html-italic">y</span>-axis) over time (<span class="html-italic">x</span>-axis). (<b>B</b>) Lineages Through Times (LTT). The number of lineages (<span class="html-italic">y</span>-axis) is displayed as a function of time (<span class="html-italic">x</span>-axis). Thin lines represent the 95% high posterior density (HPD) interval.</p>
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<p>Trend of confirmed cases of Mpox in the six WHO regions from January 2022 to June 2024. For details on the exact number of cases, see <a href="#microorganisms-12-01874-t001" class="html-table">Table 1</a>.</p>
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10 pages, 591 KiB  
Brief Report
Wastewater Surveillance of Mpox during the Summer Season of 2023 in Slovenia
by Jan Rožanec, Natalija Kranjec, Ivana Obid, Andrej Steyer, Tjaša Cerar Kišek, Tom Koritnik, Mario Fafangel and An Galičič
Infect. Dis. Rep. 2024, 16(5), 836-845; https://doi.org/10.3390/idr16050065 - 29 Aug 2024
Viewed by 627
Abstract
Since COVID-19, mpox was the first emerging pathogen to have spread globally in 2022. Wastewater-based surveillance (WBS) has proven to be an efficient early warning system for detecting potential resurgences. This report aims to provide insight into the development and implementation of WBS [...] Read more.
Since COVID-19, mpox was the first emerging pathogen to have spread globally in 2022. Wastewater-based surveillance (WBS) has proven to be an efficient early warning system for detecting potential resurgences. This report aims to provide insight into the development and implementation of WBS of mpox in Slovenia and to incorporate the surveillance results into the development of public health interventions. WBS of mpox was conducted during the period from 1 June 2023 to 30 September 2023 at the wastewater treatment plant (WWTP) Ljubljana and WWTP Koper. The selected detection method of the monkeypox virus (MPXV) in the wastewater sample was based on PCR analysis. The implemented laboratory method showed that the sample preparation and concentration method enables a stable procedure for MPXV detection in wastewater samples. The laboratory analysis of wastewater samples from the selected WWTPs did not detect the MPXV during the monitoring period. In the event of MPXV detection in a wastewater sample, targeted public health interventions would be implemented, focusing on increasing awareness among the groups of men who have sex with other men and searching for positive mpox cases. We recommend that the developed system be retained in the case of an emergency epidemiological situation. Full article
(This article belongs to the Topic Human Monkeypox Research)
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<p>Catchment areas of the wastewater treatment plant Ljubljana (green colour) and the wastewater treatment plant Koper (blue colour), included in the wastewater surveillance of mpox in Slovenia, during the summer season of 2023.</p>
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12 pages, 1520 KiB  
Article
Characterization of the Monkeypox Virus [MPX]-Specific Immune Response in MPX-Cured Individuals Using Whole Blood to Monitor Memory Response
by Elisa Petruccioli, Settimia Sbarra, Serena Vita, Andrea Salmi, Gilda Cuzzi, Patrizia De Marco, Giulia Matusali, Assunta Navarra, Luca Pierelli, Alba Grifoni, Alessandro Sette, Fabrizio Maggi, Emanuele Nicastri and Delia Goletti
Vaccines 2024, 12(9), 964; https://doi.org/10.3390/vaccines12090964 - 26 Aug 2024
Viewed by 504
Abstract
Background: Monkeypox (Mpox) is a zoonotic disease caused by monkeypox virus (MPXV), an Orthopoxvirus (OPXV). Since we are observing the first MPXV outbreak outside the African continent, the general population probably does not have a pre-existing memory response for MPXV but may have [...] Read more.
Background: Monkeypox (Mpox) is a zoonotic disease caused by monkeypox virus (MPXV), an Orthopoxvirus (OPXV). Since we are observing the first MPXV outbreak outside the African continent, the general population probably does not have a pre-existing memory response for MPXV but may have immunity against the previous smallpox vaccine based on a live replicating Vaccinia strain (VACV). Using a whole blood platform, we aim to study the MPXV- T-cell-specific response in Mpox-cured subjects. Methods: We enrolled 16 subjects diagnosed with Mpox in the previous 3–7 months and 15 healthy donors (HD) with no recent vaccination history. Whole blood was stimulated overnight with MPXV and VACV peptides to elicit CD4 and CD8 T-cell-specific responses, which were evaluated by ELISA and multiplex assay. Results: Mpox-cured subjects showed a significant IFN-γ T-cell response to MPXV and VACV. Besides IFN-γ, IL-6, IP-10, IL-8, IL-2, G-CSF, MCP-1, MIP1-α, MIP-1β, IL-1Rα, and IL-5 were significantly induced after specific stimulation compared to the unstimulated control. The specific response was mainly induced by the CD4 peptides MPX-CD4-E and VACV-CD4. Conclusions: We showed that MPXV-specific responses have a mixed Th1- and Th2-response in a whole blood platform assay, which may be useful for monitoring the specific immunity induced by vaccination or infection. Full article
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<p>MPXV-specific T-cell response from in vitro stimulated samples from Mpox-cured individuals. Graphs report the IFN-γ response induced by MPX-CD4-P; MPX-CD8-P, VACV-CD4, and VACV-CD8 stimulation. Results for the MPX-CD8-P condition were available only for 15 individuals. ELISA was performed on plasma samples from whole blood stimulation, and IFN-γ was expressed as IU/mL; the IFN-γ values of the stimulated conditions were not subtracted from the unstimulated control values. The horizontal lines represent the median; statistical analysis was performed using the Wilcoxon test; black plots refer to HIV-uninfected subjects, and red plots refer to PLWH. <b>Abbreviations:</b> IFN-γ: interferon-γ; HD: healthy donor; PLWH: people living with HIV.</p>
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<p>Multiplex analysis of immune factors different from IFN-γ specifically induced after in vitro Mpox peptide stimulation in Mpox-cured subjects. Graphs report the levels of different immune factors in response to MPX-CD4-P; MPX-CD8-P, VACV-CD4, and VACV-CD8 stimulation. The different immune factors were evaluated by Luminex assay in plasma collected after whole blood stimulation, and levels of analytes were expressed as IU/mL. The value of the stimulated condition was not subtracted from the value of the unstimulated control. Among the 27 analytes evaluated, we report only the immune factors with significant modulation between the unstimulated control and peptide stimulation. The horizontal lines represent the median; statistical analysis was performed using the Wilcoxon test; black plots refer to HIV-uninfected subjects, and red plots refer to PLWH. <b>Abbreviations:</b> IFN-γ: interferon-γ; IL: interleukin; IP-10: interferon-γ inducible protein; G-CSF: granulocyte-colony-stimulating factor; MCP-1: monocyte chemoattractant protein-1; MIP-1: monocyte chemoattractant protein-1; IL-1Ra: interleukin-1 Receptor antagonist; PLWH: people living with HIV.</p>
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<p>Mpox-specific immune signature based on selected immune factors. (<b>A</b>) The graphs represent the median proportion of immune factors secreted in response to (<b>A</b>) MPX-CD4-P; MPX-CD8-P; (<b>B</b>) VACV-CD4, VACV-CD8. The different immune factors were measured by Luminex assay in plasma collected after whole blood stimulation. The value of the stimulated condition was subtracted from the value of the unstimulated control. <b>Abbreviations:</b> IFN-γ: interferon-γ; IL: interleukin; IP-10: interferon-γ inducible protein; G-CSF: granulocyte-colony stimulating factor; MCP-1: monocyte chemoattractant protein-1; MIP-1: macrophage inflammatory protein-1; IL-1Ra: interleukin-1 Receptor antagonist.</p>
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25 pages, 9398 KiB  
Article
Modifications of Mitochondrial Network Morphology Affect the MAVS-Dependent Immune Response in L929 Murine Fibroblasts during Ectromelia Virus Infection
by Karolina Gregorczyk-Zboroch, Lidia Szulc-Dąbrowska, Pola Pruchniak, Małgorzata Gieryńska, Matylda Barbara Mielcarska, Zuzanna Biernacka, Zbigniew Wyżewski, Iwona Lasocka, Weronika Świtlik, Alicja Szepietowska, Patrycja Kukier, Aleksandra Kwiecień-Dębska and Jakub Kłęk
Pathogens 2024, 13(9), 717; https://doi.org/10.3390/pathogens13090717 - 23 Aug 2024
Viewed by 411
Abstract
Since smallpox vaccination was discontinued in 1980, there has been a resurgence of poxvirus infections, particularly the monkeypox virus. Without a global recommendation to use the smallpox vaccine, the population is not immune, posing a severe threat to public health. Given these circumstances, [...] Read more.
Since smallpox vaccination was discontinued in 1980, there has been a resurgence of poxvirus infections, particularly the monkeypox virus. Without a global recommendation to use the smallpox vaccine, the population is not immune, posing a severe threat to public health. Given these circumstances, it is crucial to understand the relationship between poxviruses and their hosts. Therefore, this study focuses on the ectromelia virus, the causative agent of mousepox, which serves as an excellent model for studying poxvirus pathogenesis. Additionally, we investigated the role of mitochondria in innate antiviral immunity during ECTV infection, focusing specifically on mitochondrial antiviral signaling protein. The study used a Moscow strain of ECTV and L929 mouse fibroblasts. Cells were treated with ECTV and chemical modulators of mitochondrial network: Mdivi-1 and CCCP. Our investigation revealed that an elongated mitochondrial network attenuates the suppression of MAVS-dependent immunity by ECTV and reduces ECTV replication in L929 fibroblasts compared to cells with an unaltered mitochondrial network. Conversely, a fragmented mitochondrial network reduces the number of progeny virions while increasing the inhibition of the virus-induced immune response during infection. In conclusion, our study showed that modifications of mitochondrial network morphology alter MAVS-dependent immunity in ECTV-infected mouse L929 fibroblasts. Full article
(This article belongs to the Special Issue Immune Response of the Host and Vaccine Development—2nd Edition)
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<p>Ectromelia virus (ECTV) replication in L929 murine fibroblasts with modified mitochondrial network morphology. The plaque assay determined the ECTV titer in L929 cells at two time points: (<b>a</b>) 18 h post infection (hpi) under various treatments, including DMSO, Mdivi-1, or CCCP, with or without transfection of poly(I:C) (MHW) LyoVec (pIC LV); and (<b>b</b>) 10 hpi with transfection of control siRNA, Mfn1 siRNA, or Drp1 siRNA or treatment with DMSO, Mdivi-1, or CCCP. The data from both (<b>a</b>,<b>b</b>) are presented as mean ± standard deviation (SD), with statistical significance indicated as * <span class="html-italic">p</span> ≤ 0.05 and ** <span class="html-italic">p</span> ≤ 0.01. ECTV suspension was obtained from cell lysates in post-culture media. Western blot analysis was performed to confirm Mfn1 or Drp1 gene silencing in target cells (<b>c</b>) and to assess ECTV antigens expression in control and infected cells treated with DMSO, Mdivi-1, or CCCP at 2, 6, 10, and 18 hpi (<b>d</b>). (<b>e</b>) Fluorescence verification of mitochondrial network morphology in L929 fibroblast after treatment with DMSO (intact network), Mdivi-1 (elongated), and CCCP (fragmented) by using MitoRed fluorochrome (red). Scale bar: 20 µm.</p>
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<p>ECTV replication in RAW 264.7 murine macrophages with modified mitochondrial network morphology. (<b>a</b>) Determination of ECTV titer by plaque assay in RAW 264.7 cells at 6 hpi treated with DMSO, Mdivi-1, or CCCP. ECTV suspension was obtained from cell lysates or post-culture media. (<b>b</b>) Fluorescence verification of mitochondrial network morphology in RAW 264.7 murine macrophages after treatment with DMSO (intact network), Mdivi-1 (elongated), and CCCP (fragmented). Macrophages were labeled using antibodies against MAVS (green) and DNA (blue). Scale bar: 20 µm.</p>
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<p>Distribution of MAVS protein in L929 cells at 18 hpi with ECTV. Cells were treated with DMSO, Mdivi-1, or CCCP. (<b>a</b>) Magnifications indicate MAVS localization in the viral factories. Arrows show the direction of fluorescence intensity measurements. Fibroblasts are labeled using antibodies against MAVS (red), HSP60 (green), or ECTV (EV; green) and DNA (blue). Scale bar: 20 µm. (<b>b</b>) Percentage of MAVS and HSP60 colocalization (<span class="html-italic">n</span> = 20). Different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) between groups.</p>
Full article ">Figure 3 Cont.
<p>Distribution of MAVS protein in L929 cells at 18 hpi with ECTV. Cells were treated with DMSO, Mdivi-1, or CCCP. (<b>a</b>) Magnifications indicate MAVS localization in the viral factories. Arrows show the direction of fluorescence intensity measurements. Fibroblasts are labeled using antibodies against MAVS (red), HSP60 (green), or ECTV (EV; green) and DNA (blue). Scale bar: 20 µm. (<b>b</b>) Percentage of MAVS and HSP60 colocalization (<span class="html-italic">n</span> = 20). Different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) between groups.</p>
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<p>Flow cytometry analysis of MAVS protein level in L929 cells at 24 hpi with ECTV (EV). (<b>a</b>) Representative histograms indicate the fluorescence intensity of APC-labeled cells treated with DMSO, Mdivi-1, or CCCP and with or without transfection with poly(I:C)(HMW) LyoVec (pIC LV). A black line is drawn through the center of the histogram of uninfected DMSO-treated cells. (<b>b</b>) Dot plots of unlabeled and APC-labeled cells treated with DMSO. (<b>c</b>) Mean fluorescence intensity (MFI) of APC in cells treated with DMSO, Mdivi-1, or CCCP and with or without transfection with poly(I:C)(HMW) LyoVec.</p>
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<p>Western blot analysis of proteins involved in MAVS-dependent immunity in L929 cells treated with DMSO, Mdivi-1, or CCCP during ECTV infection. (<b>a</b>) Representative Western blots of MDA-5, RIG-I, MAVS, pIRF3, and STING at 2, 10, 18, and 24 h post infection (hpi) with ECTV (EV). (<b>b</b>) Densitometry analysis of MDA-5, RIG-I, MAVS, pIRF3, and STING at 2, 10, 18, and 24 hpi with ECTV. The level of each protein was normalized to glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The data ARE presented as mean ± standard deviation (SD), with statistical significance indicated as * <span class="html-italic">p</span> ≤ 0.05 AND ** <span class="html-italic">p</span> ≤ 0.01. pIC LV—cells transfected with poly(I:C)(HMW) LyoVec.</p>
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<p>Oligomerization of MAVS protein in L929 cells transfected using poly(I:C)(HMW) LyoVec at 24 hpi with ECTV. (<b>a</b>,<b>b</b>) Uninfected control cells treated with Mdivi-1. Fibroblasts were labeled with specific antibodies against (<b>a</b>) MAVS (green) and pIRF3 (red) or (<b>b</b>) MAVS (red) and Hsp60 (green)—marker of mitochondria. (<b>c</b>) ECTV-infected cells treated with Mdivi-1 and labeled with specific antibodies against MAVS (red). DNA was labeled with DAPI (blue). White arrows indicate MAVS oligomers, the yellow arrow shows viral factory, and the white arrowhead indicates pIRF3 in the nucleus. Scale bar: 20 µm. (<b>d</b>) The bar chart presents the percentage of uninfected control cells with MAVS oligomers. Fibroblasts were treated with DMSO, Mdivi-1, or CCCP.</p>
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<p>Extracellular level of IFN-α (<b>a</b>) and IFN-β (<b>b</b>) produced by L929 at 24 hpi with ECTV (EV). Cells were treated using DMSO, Mdivi-1, or CCCP and/or transfected with poly(I:C)(HMW) LyoVec (pIC LV). ND—non-detected. Different letters indicate significant differences (<span class="html-italic">p</span> &lt; 0.05) between groups.</p>
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<p>Colocalization of MAVS protein with RIG-I (<b>a</b>,<b>b</b>), MDA-5 (<b>c</b>,<b>d</b>), and STING (<b>e</b>,<b>f</b>) in L929 cells at 18 hpi with ECTV. Cells were treated using DMSO, Mdivi-1, or CCCP and/or transfected with poly(I:C)(HMW) LyoVec. Box plots (<b>a</b>,<b>c</b>,<b>e</b>) indicate the number of PLA dots per cell. Different letters indicate significant differences between groups (<span class="html-italic">p</span> ≤ 0.05). (<b>b</b>,<b>d</b>,<b>f</b>) Representative figures visualize the number of PLA dots per cell in each group. Scale bar: 20 µm.</p>
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<p>Colocalization of MAVS with fission (<b>a</b>) and fusion (<b>b</b>) proteins in L929 at 18 hpi with ECTV. Cells were treated using DMSO, Mdivi-1, or CCCP and/or transfected with poly(I:C)(HMW) LyoVec. Box plots indicate the number of PLA dots per cell. Different letters indicate significant differences between groups (<span class="html-italic">p</span> ≤ 0.05).</p>
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<p>Possible interaction between MAVS and selected proteins. E3—poxviral protein; ER—endoplasmic reticulum; Drp1—dynamin-related protein 1; Fis1—Fission 1 protein; IMM—inner mitochondrial membrane; MAVS—mitochondrial antiviral signaling protein; MDA5—melanoma differentiation-associated protein 5; Mdivi-1—mitochondrial division inhibitor; Mfn1/2—mitofusin 1/2; OMM—outer mitochondrial membrane; Opa1—optic atrophy 1; RIG-I—retinoic acid-inducible gene I; STING—stimulator of interferon genes.</p>
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15 pages, 845 KiB  
Review
Assessment of the Demographic Burden of Epidemics and Diseases within the African Sub-Region: A Review of Recent Epidemiological Data, Responses, and Resultant Psychosocial Risk Tendencies
by Willy Barinem Vidona, Charity Willy-Vidona, Lekpa Kingdom David, Bassey Monday Abia, John Nwolim Paul and Ukpai Agwu Eze
Hygiene 2024, 4(3), 346-360; https://doi.org/10.3390/hygiene4030027 - 22 Aug 2024
Viewed by 720
Abstract
Background: Over the past few decades, epidemic outbreaks and disease occurrences have become more frequent and widespread in Africa, posing challenges for poor countries in the region and impacting vulnerable populations. Limited resources, inadequate healthcare infrastructure, socioeconomic inequalities, malnutrition, mass gatherings, and lack [...] Read more.
Background: Over the past few decades, epidemic outbreaks and disease occurrences have become more frequent and widespread in Africa, posing challenges for poor countries in the region and impacting vulnerable populations. Limited resources, inadequate healthcare infrastructure, socioeconomic inequalities, malnutrition, mass gatherings, and lack of proper sanitation and hygiene contribute to Africa’s high burden of epidemics and infectious diseases such as cholera, measles, monkeypox, Ebola viral disease, and COVID-19. This paper reviewed epidemic outbreaks and diseases transmitted through intimate contact in Africa and their impacts on vulnerable populations. Methods: A narrative review approach was adopted to gather and analyze the relevant literature on epidemic outbreaks in African sub-regions. The review encompassed causes, transmission patterns, demographic impact, community susceptibility, preventive measures, and psychosocial risk-taking behaviors. Results: This study emphasizes the importance of addressing the causes of response lapses and the resulting human, material, and economic losses in the region. Effective preventive measures include disease surveillance, early mitigation strategies, contact tracing, personal protective measures, improved political and public health leadership, and socioeconomic equity. However, the review highlights challenges in implementing these measures effectively due to limited resources, delayed detection, and difficulties in scaling up response solutions. Conclusions: There is a need for a comprehensive approach involving health departments, infrastructure development, and addressing socioeconomic factors. Full article
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<p>The most reported epidemic outbreaks in Africa as of December 2023. Data extracted from Africa Centre for Disease Control and Prevention, Africa CDC Epidemic Intelligence Report, 29 March 2024 [<a href="#B5-hygiene-04-00027" class="html-bibr">5</a>].</p>
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<p>The most reported epidemic outbreaks in Africa as of March 2024. Data extracted from Africa Centre for Disease Control and Prevention, Epidemic Intelligence Report, 29 March 2024 [<a href="#B5-hygiene-04-00027" class="html-bibr">5</a>].</p>
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<p>Cumulative number of COVID-19 cases and deaths reported to WHO from January 2020 to 7 July 2024. Source: WHO 2024 [<a href="#B18-hygiene-04-00027" class="html-bibr">18</a>].</p>
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9 pages, 589 KiB  
Article
Monkeypox in Bulgaria: Significance of Various Clinical Samples, Clinical Manifestation, and Molecular Detection
by Stefka Krumova, Radostina Stefanova, Petia Genova-Kalou, Daniel Ivanov, Maria Pishmisheva, Stanislav Kotsev and Iva Christova
J. Clin. Med. 2024, 13(16), 4856; https://doi.org/10.3390/jcm13164856 - 17 Aug 2024
Viewed by 1498
Abstract
Background/Objectives: Monkeypox (mpox) is currently the most common orthopoxvirus (OPXV) zoonotic disease, and, since 2022, there has been atypical person-to-person transmission observed in non-endemic countries. The present study aimed to investigate the frequency of monkeypox virus (MPXV) and OPXV DNA detection in [...] Read more.
Background/Objectives: Monkeypox (mpox) is currently the most common orthopoxvirus (OPXV) zoonotic disease, and, since 2022, there has been atypical person-to-person transmission observed in non-endemic countries. The present study aimed to investigate the frequency of monkeypox virus (MPXV) and OPXV DNA detection in recommended and alternative clinical materials taken during the acute and convalescent phases of infection in Bulgarian patients. Methods: The study included laboratory investigation by real time PCR of 181 clinical samples from 42 Bulgarian patients with possible mpox infections. Results: MPXV DNA was detected in 23/181 (12.71%), and OPXV DNA in 20/181 (11.05%) clinical samples. There were six mpox-confirmed patients aged 23 to 44. At the highest frequency, MPXV and OPXV DNA were detected in samples of vesicular contents (6/6) and nasal/oropharyngeal secretions (5/6 and 4/6) during the first three days from the appearance of clinical symptoms. We demonstrated MPXV and OPXV DNA in alternative samples (urine, feces, ejaculate, and saliva), and in follow-up patient samples, taken two weeks after mpox confirmation in the convalescent phase (vesicular contentsand urine). Conclusions: Our findings suggested that MPXV may be detected in a larger set of clinical materials, including alternatives, where the virus can persist for more than two weeks. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
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<p>Evidence of MPXV DNA in the study of different types of clinical material samples taken during the acute and convalescent phase of infection in Bulgarian patients.</p>
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19 pages, 511 KiB  
Article
Modeling and Analysis of Monkeypox Outbreak Using a New Time Series Ensemble Technique
by Wilfredo Meza Cuba, Juan Carlos Huaman Alfaro, Hasnain Iftikhar and Javier Linkolk López-Gonzales
Axioms 2024, 13(8), 554; https://doi.org/10.3390/axioms13080554 - 14 Aug 2024
Cited by 1 | Viewed by 897
Abstract
The coronavirus pandemic has raised concerns about the emergence of other viral infections, such as monkeypox, which has become a significant hazard to public health. Thus, this work proposes a novel time series ensemble technique for analyzing and forecasting the spread of monkeypox [...] Read more.
The coronavirus pandemic has raised concerns about the emergence of other viral infections, such as monkeypox, which has become a significant hazard to public health. Thus, this work proposes a novel time series ensemble technique for analyzing and forecasting the spread of monkeypox in the four highly infected countries with the monkeypox virus. This approach involved processing the first cumulative confirmed case time series to address variance stabilization, normalization, stationarity, and a nonlinear secular trend component. After that, five single time series models and three proposed ensemble models are used to estimate the filtered confirmed case time series. The accuracy of the models is evaluated using typical accuracy mean errors, graphical evaluation, and an equal forecasting accuracy statistical test. Based on the results, it is found that the proposed time series ensemble forecasting approach is an efficient and accurate way to forecast the cumulative confirmed cases for the top four countries in the world and the entire world. Using the best ensemble model, a forecast is made for the next 28 days (four weeks), which will help understand the spread of the disease and the associated risks. This information can prevent further spread and enable timely and effective treatment. Furthermore, the developed novel time series ensemble approach can be used to forecast other diseases in the future. Full article
(This article belongs to the Special Issue Modeling and Analysis of Complex Network)
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<p>Monkeypox infections forecasting: A complete proposed time series ensemble approach layout.</p>
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<p>Comparison of daily confirmed monkeypox cases in four most affected countries from 1 June 2022 to 31 July 2023.</p>
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<p>Comparison of daily cumulative confirmed monkeypox cases with superimposed the nonlinear trend component in four most affected countries from 1 June 2022 to 31 July 2023.</p>
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<p>Residual series after extracting the nonlinear trend component in the four most affected countries and the entire world case within the proposed forecasting technique.</p>
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11 pages, 801 KiB  
Article
A Comparative Evaluation of Three Diagnostic Assays for the Detection of Human Monkeypox
by Jing Qu, Xiaomin Zhang, Kun Liu, You Li, Ting Wang, Zhonggang Fang, Cheng Chen, Xiao Tan, Ying Lin, Qing Xu, Yan Yang, Wanqing Wang, Manyu Huang, Shiliang Guo, Ziqiu Chen, Wei Rao, Xiaolu Shi and Bo Peng
Viruses 2024, 16(8), 1286; https://doi.org/10.3390/v16081286 - 12 Aug 2024
Cited by 1 | Viewed by 1418
Abstract
Accurate and early diagnosis of monkeypox virus (MPXV) is crucial for controlling epidemics and treating affected individuals promptly. This study aimed to assess the analytical and clinical performance of the MolecisionTM Monkeypox Virus qPCR Assay, Biorain Monkeypox Virus ddPCR Assay, and MAGLUMI [...] Read more.
Accurate and early diagnosis of monkeypox virus (MPXV) is crucial for controlling epidemics and treating affected individuals promptly. This study aimed to assess the analytical and clinical performance of the MolecisionTM Monkeypox Virus qPCR Assay, Biorain Monkeypox Virus ddPCR Assay, and MAGLUMI® Monkeypox Virus Ag (chemiluminescence immunoassay, CLIA) Assay. Additionally, it aimed to compare the clinical application of antigen and nucleic acid assays to offer insights into using commercial monkeypox assay kits. Specimens from 117 clinical patients, serial diluted virus cell culture supernatant, and artificially created positive samples were tested to evaluate the performance of these assay kits for MPXV diagnostics. The Biorain Monkeypox Virus ddPCR Assay had a limit of detection (LoD) of 3.89 CCID50/mL, while the MolecisionTM Monkeypox Virus qPCR Assay had an LoD of 15.55 CCID50/mL. The MAGLUMI® Monkeypox Virus Ag (CLIA) Assay had an LoD of 0.500 pg/mL. The accuracy of the MolecisionTM Monkeypox Virus qPCR Assay was comparable to the Biorain Monkeypox Virus ddPCR Assay, and the MAGLUMI® Monkeypox Virus Ag (CLIA) Assay demonstrated high sensitivity. The specificity of all three MPXV diagnostic assays for clinical specimens with potential cross-reacting substances was 100%. In conclusion, this study provides valuable insights into the clinical application of monkeypox assays, supporting efforts to mitigate and control the spread of monkeypox. Full article
(This article belongs to the Topic Human Monkeypox Research)
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Graphical abstract

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<p>Relative sensitivity of MAGLUMI<sup>®</sup> Monkeypox Virus Ag (CLIA) Assay in qPCR assay Ct intervals. Each blue square within the illustration signifies the sensitivity of CLIA assay, accompanied by 95% confidence intervals denoted by vertical lines. These values are presented in relation to specific qPCR Ct intervals, including &lt;29, &lt;30, &lt;31, &lt;32, &lt;32, &lt;33, &lt;34, &lt;35, and &lt;40.</p>
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<p>The correlation of MPXV DNA measures from the Molecision<sup>TM</sup> Monkeypox Virus qPCR Assay/Biorain Monkeypox Virus ddPCR Assay. Scatter plots with a regression line (red line) for the Molecision<sup>TM</sup> Monkeypox Virus qPCR Assay and Biorain Monkeypox Virus ddPCR Assay. Each black dot symbolizes a specimen both with results from both qPCR assay and ddPCR assay. The results of the ddPCR assay were log-transformed as log<sub>10</sub> (number of copies/mL) for the purpose of simple linear regression.</p>
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21 pages, 4390 KiB  
Article
A Combined Transcriptomic and Proteomic Analysis of Monkeypox Virus A23 Protein on HEK293T Cells
by Yihao Wang, Yihan Li, Mingzhi Li, Keyi Wang, Jiaqi Xiong, Ting Wang, Yu Wang, Yunli Guo, Lingbao Kong and Meifeng Li
Int. J. Mol. Sci. 2024, 25(16), 8678; https://doi.org/10.3390/ijms25168678 - 8 Aug 2024
Viewed by 856
Abstract
Monkeypox virus (MPXV) is a cross-kingdom pathogen infecting both humans and wildlife, which poses a significant health risk to the public. Although MPXV attracts broad attention, there is a lack of adequate studies to elucidate pathogenic mechanisms associated with viral infections. In this [...] Read more.
Monkeypox virus (MPXV) is a cross-kingdom pathogen infecting both humans and wildlife, which poses a significant health risk to the public. Although MPXV attracts broad attention, there is a lack of adequate studies to elucidate pathogenic mechanisms associated with viral infections. In this study, a high-throughput RNA sequencing (RNA-seq) and liquid chromatography–tandem mass spectrometry (LC-MS/MS) approach was used to explore the transcriptional and metabolic responses of MPXV A23 protein to HEK293T cells. The protein–protein interactions and signaling pathways were conducted by GO and KEGG analyses. The localization of A23 protein in HEK293T cells was detected by immunofluorescence. A total of 648 differentially expressed genes (DEGs) were identified in cells by RNA-Seq, including 314 upregulated genes and 334 downregulated genes. Additionally, liquid chromatography–tandem mass spectrometry (LC-MS/MS) detected 115 cellular proteins that interact with the A23 proteins. Transcriptomic sequencing analysis revealed that transfection of MPXV A23 protein modulated genes primarily associated with cellular apoptosis and DNA damage repair. Proteomic analysis indicated that this protein primarily interacted with host ribosomal proteins and histones. Following the identification of the nuclear localization sequence RKKR within the A23 protein, a truncated mutant A23ΔRKKR was constructed to investigate the subcellular localization of A23 protein. The wild-type A23 protein exhibits a significantly higher nuclear-to-cytoplasmic ratio, exceeding 1.5, in contrast to the mutant A23ΔRKKR, which has a ratio of approximately 1. Immunofluorescence assays showed that the A23 protein was mainly localized in the nucleus. The integration of transcriptomics and proteomics analysis provides a comprehensive understanding of the interaction between MPXV A23 protein and the host. Our findings highlight the potential role of this enzyme in suppressing host antiviral immune responses and modulating host gene expression. Full article
(This article belongs to the Special Issue Recent Advances in Herpesviruses)
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<p>Construction and expression of recombinant A23R in HEK293T cells. (<b>A</b>) Model of a cloned fragment. (<b>B</b>) Double digestion of the pCAGGS−HA−A23R plasmid. Lane 1–4: A23R recombinants were digested by <span class="html-italic">EcoR</span> I and <span class="html-italic">Xho</span> I. (<b>C</b>) Expression of recombinant A23R in HEK293T cells, followed by immunoblot analysis using HA−tag antibodies or anti β−actin antibodies. Lane 1: Transfected with pCAGGS−HA. Lane 2: Transfected with pCAGGS−HA−A23R. (<b>D</b>) Western blot analysis of the Co−IP sample. Flow Through: the WCL after anti−HA incubate.</p>
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<p>Construction and expression of recombinant A23R in HEK293T cells. (<b>A</b>) Model of a cloned fragment. (<b>B</b>) Double digestion of the pCAGGS−HA−A23R plasmid. Lane 1–4: A23R recombinants were digested by <span class="html-italic">EcoR</span> I and <span class="html-italic">Xho</span> I. (<b>C</b>) Expression of recombinant A23R in HEK293T cells, followed by immunoblot analysis using HA−tag antibodies or anti β−actin antibodies. Lane 1: Transfected with pCAGGS−HA. Lane 2: Transfected with pCAGGS−HA−A23R. (<b>D</b>) Western blot analysis of the Co−IP sample. Flow Through: the WCL after anti−HA incubate.</p>
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<p>Identification of differentially expressed genes (DEGs). (<b>A</b>) The volcano diagram of DEGs. The horizontal axis represents the fold change in gene expression between the experimental and control groups (log<sub>2</sub>FoldChange). The vertical axis represents the significance of the DEGs between the experimental and control groups (−log<sub>10</sub>padj or −log<sub>10</sub>pvalue). Up−regulated genes are shown as red dots. Down−regulated genes are shown as green dots. Blue dots indicate no statistically significant genes (NO 25029). Threshold lines for DEGs screening criteria are indicated by blue dashed lines. (<b>B</b>) Heatmap of DEGs with length, type, and chr. The horizontal coordinate represents the sample name. The vertical coordinates on the left represent the cluster analysis. The vertical coordinates on the right represent length/type/chr. The heatmap specifies the length of each gene (length), categorizes its functions (type), and determines its position in the chromosome (chr). The red color in the middle of the heatmap represents high expression, and the green color represents low expression.</p>
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<p>Identification of differentially expressed genes (DEGs). (<b>A</b>) The volcano diagram of DEGs. The horizontal axis represents the fold change in gene expression between the experimental and control groups (log<sub>2</sub>FoldChange). The vertical axis represents the significance of the DEGs between the experimental and control groups (−log<sub>10</sub>padj or −log<sub>10</sub>pvalue). Up−regulated genes are shown as red dots. Down−regulated genes are shown as green dots. Blue dots indicate no statistically significant genes (NO 25029). Threshold lines for DEGs screening criteria are indicated by blue dashed lines. (<b>B</b>) Heatmap of DEGs with length, type, and chr. The horizontal coordinate represents the sample name. The vertical coordinates on the left represent the cluster analysis. The vertical coordinates on the right represent length/type/chr. The heatmap specifies the length of each gene (length), categorizes its functions (type), and determines its position in the chromosome (chr). The red color in the middle of the heatmap represents high expression, and the green color represents low expression.</p>
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<p>RT−qPCR verified the expression of seven genes. We examined the gene expression levels of <span class="html-italic">IL9R, PLA2G4C, MAFA, CYP2E1, H3C1, H2BC17, HLA−DPB1 LIG4, BIRC3, SMAC, RAD9</span>, and <span class="html-italic">SGO2</span> by RT−qPCR in HEK293T cells after the expression of A23 protein. RNA expression levels in each system were normalized to β−actin. The error bars indicate the SD of repeated RT−qPCR. All experiments were conducted in−dependently, at least three times. Statistical significance is indicated by ** <span class="html-italic">p</span> &lt; 0.01, *** <span class="html-italic">p</span> &lt; 0.001, **** <span class="html-italic">p</span> &lt; 0.0001.</p>
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<p>Functional enrichment analysis of DEGs. The scatter plot of GO (<b>A</b>) and KEGG (<b>B</b>) analysis. The vertical axis represents the top thirty terms with the most significance. The horizontal axis represents the gene ratio. Count: the number of DEGs. Gene ratio: the ratio of DEG number to background gene number. <span class="html-italic">p</span>-value: indicators of the significance of the term; the smaller the <span class="html-italic">p</span>-value, the more significant the term.</p>
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<p>Functional enrichment analysis of the interacting proteins. The scatter plot of GO (<b>A</b>) and KEGG (<b>B</b>) analysis. The vertical axis represents the top thirty terms with the most significance. The horizontal axis represents the ratio. Count: the number of proteins. Gene ratio: the ratio of protein number to background protein number. <span class="html-italic">p</span>-value: indicators of the significance of the term, the smaller the <span class="html-italic">p</span>-value, the more significant the term.</p>
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<p>Functional enrichment analysis of the interacting proteins. The scatter plot of GO (<b>A</b>) and KEGG (<b>B</b>) analysis. The vertical axis represents the top thirty terms with the most significance. The horizontal axis represents the ratio. Count: the number of proteins. Gene ratio: the ratio of protein number to background protein number. <span class="html-italic">p</span>-value: indicators of the significance of the term, the smaller the <span class="html-italic">p</span>-value, the more significant the term.</p>
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<p>Mutation of the RKKR impairs nuclear import of A23 protein. The nuclear location signal in A23 protein was identified through NLStradamus (<b>A</b>), NucPred (<b>B</b>), and cNLS Mapper (<b>C</b>). HEK293T cells were transiently transfected with plasmic coding for HA-A23 and the A23<sub>△RKKR</sub>. (<b>D</b>) Transiently transfected cells were fixed, stained with DAPI, and analyzed by fluorescent Inverted microscope. (<b>E</b>) Nuclear localization of A23 protein and mutants was assessed in transiently transfected cells as a ratio of nuclear to cytoplasmic fluorescence using the Image J software version 1.54j. Data are from n = 4 fluorescent cells analyzed. ** <span class="html-italic">p</span> &lt; 0.05 difference from WT-A23 transfected cells. Scale bar = 50 µm.</p>
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<p>Mutation of the RKKR impairs nuclear import of A23 protein. The nuclear location signal in A23 protein was identified through NLStradamus (<b>A</b>), NucPred (<b>B</b>), and cNLS Mapper (<b>C</b>). HEK293T cells were transiently transfected with plasmic coding for HA-A23 and the A23<sub>△RKKR</sub>. (<b>D</b>) Transiently transfected cells were fixed, stained with DAPI, and analyzed by fluorescent Inverted microscope. (<b>E</b>) Nuclear localization of A23 protein and mutants was assessed in transiently transfected cells as a ratio of nuclear to cytoplasmic fluorescence using the Image J software version 1.54j. Data are from n = 4 fluorescent cells analyzed. ** <span class="html-italic">p</span> &lt; 0.05 difference from WT-A23 transfected cells. Scale bar = 50 µm.</p>
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<p>A combined transcriptomic and proteomic analysis of monkeypox virus A23 protein on HEK293T cells.</p>
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13 pages, 4010 KiB  
Article
Characterization of Human Immortalized Keratinocyte Cells Infected by Monkeypox Virus
by Chaode Gu, Zhiqiang Huang, Yongyang Sun, Shaowen Shi, Xiubo Li, Nan Li, Yang Liu, Zhendong Guo, Ningyi Jin, Zongzheng Zhao, Xiao Li and Hongwei Wang
Viruses 2024, 16(8), 1206; https://doi.org/10.3390/v16081206 - 26 Jul 2024
Viewed by 733
Abstract
Monkeypox virus (MPXV) can induce systemic skin lesions after infection. This research focused on studying MPXV proliferation and the response of keratinocytes. Using transmission electron microscopy (TEM), we visualized different stages of MPXV development in human immortalized keratinocytes (HaCaT). We identified exocytosis of [...] Read more.
Monkeypox virus (MPXV) can induce systemic skin lesions after infection. This research focused on studying MPXV proliferation and the response of keratinocytes. Using transmission electron microscopy (TEM), we visualized different stages of MPXV development in human immortalized keratinocytes (HaCaT). We identified exocytosis of enveloped viruses as the exit mechanism for MPXV in HaCaT cells. Infected keratinocytes showed submicroscopic changes, such as the formation of vesicle-like structures through the recombination of rough endoplasmic reticulum membranes and alterations in mitochondrial morphology. Transcriptome analysis revealed the suppressed genes related to interferon pathway activation and the reduced expression of antimicrobial peptides and chemokines, which may facilitate viral immune evasion. In addition, pathway enrichment analysis highlighted systemic lupus erythematosus pathway activation and the inhibition of the Toll-like receptor signaling and retinol metabolism pathways, providing insights into the mechanisms underlying MPXV-induced skin lesions. This study advances our understanding of MPXV’s interaction with keratinocytes and the complex mechanisms leading to skin lesions. Full article
(This article belongs to the Section General Virology)
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<p>The phylogenetic analysis of hMpxV/China/GZ8H-01/2023. Phylogenetic analysis showed the hMpxV/China/GZ8H-01/2023 isolate belonged to clade IIb of MPXV, highlighted in red.</p>
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<p>The growth dynamics of MPXV in HaCaT cells. (<b>A</b>) Cytopathic effects (CPE) of HaCaT cells infected with MPXV by crystal violet staining. (<b>B</b>) CPE of HaCaT cells infected with 0.5 MOI MPXV at different time points (0, 24, 48, 72, and 96 hpi). (<b>C</b>) HaCaT cells infected with 0.1 or 0.5 MOI MPXV, virus concentration in the supernatant were detected and recorded at 0, 24, 48, 72, and 96 hpi, respectively (<span class="html-italic">n</span> = 3). (<b>D</b>) Infection of Vero-E6 and HaCaT cells with 0.5 MOI MPXV, followed by virus concentration in the supernatant detection and recording at 24 and 48 hpi, respectively (<span class="html-italic">n</span> = 3). Data were analyzed by one-way ANOVA and <span class="html-italic">t</span>-test. <span class="html-italic">p</span>  &lt;  0.05 was considered statistically significant. “***” represents <span class="html-italic">p</span>  &lt;  0.001.</p>
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<p>Morphogenesis of MPXV in HaCaT cells. (<b>A</b>) Numerous MPXV virions were observed in cytoplasmic matrix of the infected HaCaT cells. (<b>B</b>,<b>C</b>) The mature MPXV virions presented two different forms, the unencapsulated virion IMV and the encapsulated virion IEV. (<b>D</b>) The viral factory (marked with a yellow pentagram) encompassed various structures, including crescent structure (green triangle), immature (blue triangle) and mature (red triangle) viral particles, as well as multi-membrane vesicles (green asterisk), and abnormal mitochondria (blue asterisk). (<b>E</b>) Transformation of MPXV from viral crescent structures to IMV. (<b>F</b>) The mature virions were enveloped by membranes. (<b>G</b>) IEV at the periphery of the cell. (<b>H</b>) IEV was transported out of the cell through exocytosis. (<b>I</b>) The extracellular virions enveloped by singular membrane near the plasma membrane were termed as CEV. (<b>J</b>) Released extracellular virions were termed as EEV.</p>
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<p>Ultrastructural features of HaCaT cells infected with MPXV. (<b>A</b>) The HaCaT cells at the early stage of MPXV infection exhibited a substantial presence of EEV attached to the cell membrane surface, along with a few virions within the envelope, while maintaining an intact mitochondrial structure. (<b>B</b>) Mitochondria in late-stage infected cells showed an enlarged and rounded morphology, accompanied by a reduction in or the disappearance of cristae, with the matrices filled with granular contents. (<b>C</b>) Different forms of spherical mitochondria showed varying degrees of damage to their inner and outer membranes or cristae. (<b>D</b>) Numerous vesicle structures derived from rER were observed in the infected HaCaT cells. (<b>E</b>) The moment when the rER membrane protruded outward to form a vesicle. (<b>F</b>) At the late stage of MPXV infection, abundant multi-membrane vesicle structures appeared in HaCaT cells, along with the presence of mature virions nearby. (<b>G</b>) The nucleus in a portion of infected HaCaT cells exhibited a C-type morphology, with an opening facing the region of MPXV aggregation.</p>
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<p>Ultrastructural features of HaCaT cells infected with MPXV. (<b>A</b>) The HaCaT cells at the early stage of MPXV infection exhibited a substantial presence of EEV attached to the cell membrane surface, along with a few virions within the envelope, while maintaining an intact mitochondrial structure. (<b>B</b>) Mitochondria in late-stage infected cells showed an enlarged and rounded morphology, accompanied by a reduction in or the disappearance of cristae, with the matrices filled with granular contents. (<b>C</b>) Different forms of spherical mitochondria showed varying degrees of damage to their inner and outer membranes or cristae. (<b>D</b>) Numerous vesicle structures derived from rER were observed in the infected HaCaT cells. (<b>E</b>) The moment when the rER membrane protruded outward to form a vesicle. (<b>F</b>) At the late stage of MPXV infection, abundant multi-membrane vesicle structures appeared in HaCaT cells, along with the presence of mature virions nearby. (<b>G</b>) The nucleus in a portion of infected HaCaT cells exhibited a C-type morphology, with an opening facing the region of MPXV aggregation.</p>
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<p>RNA-seq analyses of the MPXV-infected HaCaT cells. (<b>A</b>) The location of MPXV transcript in viral genome. (<b>B</b>) The volcano plot of DEGs regarding uninfected and 24 h post-infected HaCaT cells (Log2 (fold change) &gt; 1, adjusted <span class="html-italic">p</span> &lt; 0.05). The red dots represent upregulated genes while the blue dots represent downregulated genes. (<b>C</b>) The heat map of DEGs, in which the genes associated with interferon and IL17 signaling pathways were marked. (<b>D</b>) KEGG analysis of the DEGs. (<b>E</b>) DEGs were categorized based on their gene ontology (GO) terms related to biological process (BP), cell component (CC), and molecular function (MF). (<b>F</b>) Gene set enrichment analysis of specified gene sets, including lupus erythematosus signaling pathway, body assembly, Toll-like receptor signaling pathway, and retinoid metabolism.</p>
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9 pages, 2647 KiB  
Brief Report
The Generation and Characterization of Monoclonal Antibodies against the MPXV A29L Protein
by Wenlong Zhu, Mengjia Zhang, Mengdi Zhang, Ran Jing, Jiaru Zhou, Hua Cao, Changcheng Liu, Hongmei Zhu, Ahmed H. Ghonaim, Sherin R. Rouby and Wentao Li
Viruses 2024, 16(8), 1184; https://doi.org/10.3390/v16081184 - 24 Jul 2024
Viewed by 919
Abstract
Mpox (formerly known as monkeypox) is a zoonotic disease caused by monkeypox virus (MPXV), a DNA virus belonging to the Orthopoxvirus genus, in the Poxviridae family. The disease constitutes a moderate risk to public health at the global level. The MPXV A29L protein [...] Read more.
Mpox (formerly known as monkeypox) is a zoonotic disease caused by monkeypox virus (MPXV), a DNA virus belonging to the Orthopoxvirus genus, in the Poxviridae family. The disease constitutes a moderate risk to public health at the global level. The MPXV A29L protein plays a crucial role in coordinating virion assembly and facilitating important virus-host interactions. This study focused on the expression, purification, and recombinant protein synthesis of the A29L protein of MPXV using prokaryotic systems. Using hybridoma technology, we successfully generated the monoclonal antibodies (mAbs) 1E12 and 4B2, which specifically recognize the A29L protein. These mAbs were found to be suitable for use in indirect immunofluorescence assays (IFA), Western blotting, and immunoprecipitation (IP). Our investigation also revealed that mAbs 1E12 and 4B2 could detect the A27L protein, a homologous protein found in the vaccinia virus Western Reserve (VACV WR) strain, using IFA, Western blotting, and immunoprecipitation (IP). Using mAbs 1E12 and 4B2 as primary immunological probes, A27L protein expression was detected as early as 6 h postinfection with VACV WR, with increasing protein levels being observed throughout the infection. This study enhances our understanding of the protein structure and function of MPXV and contributes to the development of specific MPXV detection methods. Full article
(This article belongs to the Special Issue B Cell-Mediated Immunity to Viruses)
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<p>The expression and purification of the recombinant A29L protein and purification of the mAbs. (<b>A</b>) Lane 1: amplified products of the MPXV A29L gene. Lane 2: linearized vectors obtained by PCR amplification. (<b>B</b>) The purification of the recombinant A29L protein identified by SDS-PAGE. (<b>C</b>) A Western blot analysis of the recombinant A29L protein using an anti-His mAb. (<b>D</b>) The identification of the purified mAbs 1E12 and 4B2.</p>
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<p>The specificity and immunogenicity of the mAbs 1E12 and 4B2 against the MPXV A29L protein strain were determined by Western blotting and IFA. (<b>A</b>) The Western blot detection of the A29L protein’s overexpression in HEK-293T cells using mAb 1E12, mAb 4B2, and anti-Flag mAb. (<b>B</b>) The IFA results of mAb 1E12 and mAb 4B2 in HEK-293T cells transfected with pCAGGS-A29L. (<b>C</b>) The lysates of HEK-293T cells were immunoprecipitated with either mAb 1E12 (left) or mAb 4B2 (right). The precipitates were analyzed by Western blotting using antibodies against Flag.</p>
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<p>The cross-reactivity of mAbs 1E12 and 4B2 against the A27L protein of VACV. (<b>A</b>) A comparison of the amino acid sequence similarity between MPXV A29L and VACV WR A27L. (<b>B</b>) The Western blot detection of mAbs. The reactivity of mAb 1E12 and mAb 4B2 with the VACV WR A27L protein in infected cells was analyzed by Western blotting. (<b>C</b>) The IFA detection of mAb 1E12 and mAb 4B2 in VACV WR-infected cells. The green fluorescence represents the reaction of mAb 1E12 and mAb 4B2 with A27L in different strains of VACV. (<b>D</b>) The detection of the IP capacities of mAb 1E12 and mAb 4B2. The VACV WR-infected cells were lysed and immunoprecipitated with either mAb 1E12 or mAb 4B2. The precipitates were analyzed by Western blotting using antibodies against A27L.</p>
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<p>The detection of A27L protein expression at different time points after VACV infection. The expression of the A27L protein in VACV WR-infected cells was monitored at various time points postinfection using IFA. Scale bar, 50 μm.</p>
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10 pages, 857 KiB  
Case Report
Primary Varicella Infection in a Young Adult from the Democratic Republic of the Congo: A Case Report and Mini-Review
by Andrew McNaughton, Nessika Karsenti, Jason Kwan, Asal Adawi, Saniya Mansuri and Andrea K. Boggild
Infect. Dis. Rep. 2024, 16(4), 628-637; https://doi.org/10.3390/idr16040048 - 19 Jul 2024
Viewed by 777
Abstract
We describe a case of an immunocompetent adult male patient originally from the Democratic Republic of Congo (DRC), who was referred to our unit for a several-day history of fever and a pruritic, vesicular rash. There was initial concern in the Emergency Department [...] Read more.
We describe a case of an immunocompetent adult male patient originally from the Democratic Republic of Congo (DRC), who was referred to our unit for a several-day history of fever and a pruritic, vesicular rash. There was initial concern in the Emergency Department for Mpox (formerly known as “monkeypox”) given the current epidemiology versus other viral etiologies. Primary varicella zoster virus (pVZV) infection was ultimately diagnosed by PCR from a swabbed, unroofed lesion, and he recovered completely with supportive management and without antiviral therapy. We herein describe how common viral exanthems may best be differentiated in an emergency or outpatient setting. Full article
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<p>Umbilicated and vesicular rash on Days 4–5 of symptoms. (<b>A</b>)—Backside of patient on Day 4, during his emergency department visit. (<b>B</b>)—Right thigh of patient on Day 5, one day prior to consultation with our unit. (<b>C</b>)—Left arm of patient on Day 5 of illness. (<b>D</b>)—Close up of right thigh on Day 5 of illness.</p>
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<p>Timeline depicting the onset and progression of symptoms and events leading to diagnosis and resolution.</p>
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7 pages, 214 KiB  
Viewpoint
High Rates of Miscarriage and Stillbirth among Pregnant Women with Clade I Mpox (Monkeypox) Are Confirmed during 2023–2024 DR Congo Outbreak in South Kivu Province
by David A. Schwartz
Viruses 2024, 16(7), 1123; https://doi.org/10.3390/v16071123 - 13 Jul 2024
Cited by 1 | Viewed by 1756
Abstract
Mpox (monkeypox) is a neglected tropical disease that has received increased attention since the multi-nation outbreak that began in 2022. The virus is endemic in West and Central Africa, where the Democratic Republic of the Congo (DRC) is the most affected country. Clade [...] Read more.
Mpox (monkeypox) is a neglected tropical disease that has received increased attention since the multi-nation outbreak that began in 2022. The virus is endemic in West and Central Africa, where the Democratic Republic of the Congo (DRC) is the most affected country. Clade I monkeypox virus (MPXV) infection is endemic in the DRC and has an overall case fatality rate of 10.6% among children and adults. A study conducted in Sankuru Province, DRC, from 2007 to 2011 demonstrated that 75% of pregnant women with mpox had miscarriages or stillbirth. Further analysis of a stillborn fetus showed that MPXV could infect both the placenta and fetus, causing congenital infection. No additional cases of Clade I MPXV in pregnant women were reported until a new outbreak occurred in South Kivu Province during 2023 and 2024. Eight pregnant women having Clade I MPXV infection were identified, of whom four had either miscarriages or stillbirth, representing a 50% fetal mortality rate. These reports confirm previous data from the DRC that indicate the capability of Clade I MPXV to affect the fetus, causing congenital infection and fetal loss in a high percentage of cases. In this article, we review both past and new data from the DRC on the effects of Clade I MPXV during pregnancy and discuss the association of mpox with fetal loss. Full article
20 pages, 1120 KiB  
Article
A Framework for Messy Communication: A Qualitative Study of Competing Voices of Authority on Social Media
by Seth J. Meyer and Kimberly Wiley
Adm. Sci. 2024, 14(7), 145; https://doi.org/10.3390/admsci14070145 - 8 Jul 2024
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Abstract
While governments, nonprofits, and influencers differ dramatically in their resource availability for social media content creation and their duties to public safety, health, and welfare, all play a role in communication with the public at large. Governments provide for the broader public good [...] Read more.
While governments, nonprofits, and influencers differ dramatically in their resource availability for social media content creation and their duties to public safety, health, and welfare, all play a role in communication with the public at large. Governments provide for the broader public good and nonprofits and influencers have the opportunity to serve isolated communities vulnerable to social ills and health crises. We explore how these three content creator groups concurrently and independently use social media to provide critical information. We qualitatively coded 1392 posts by governments, nonprofits, and influencers on 6 social media platforms shared across the United States during the 2022 Mpox outbreak. We constructed a framework defining a public health communication progression from chaotic to controlled. The findings indicated that governments can reach minoritized communities to resolve a public health crisis by partnering with nonprofits and influencers trusted by these audiences and, most importantly, practicing flexible control over shared messaging. Full article
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Figure 1

Figure 1
<p>A framework for detangling public administrations’ messy social media communication.</p>
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