Monkeypox-related ophthalmic disease (MPXROD): Monitoring the antiviral effect of tecovirimat with monkeypox virus detection in tear samples.

INTRODUCTION: Although monkeypox-related ophthalmic disease (MPXROD) is rare, visual impairing complications have been reported. At present, tecovirimat is the standard-of-care antiviral treatment. In this MPXROD case, the effect of tecovirimat was assessed with PCR analysis of tear samples and concurrent monitoring of inflammation with laser flare photometry (LFP). CASE DESCRIPTION: The patient presented with a palpebral lesion and a corneal ulcer in his right eye, with complete absence of the corneal epithelium, high intraocular pressure and anterior uveitis. MPXV-DNA was detected in tear samples with real-time PCR (RT-PCR). A total volume of 0.5 ml tear-wash was aspirated from the inferior fornix, following instillation of saline onto the ocular surface. In addition, LFP was used to quantify inflammation in both eyes. Viral load in tear samples was detected prior to treatment initiation. In the left eye, tear samples tested negative for MPXV-DNA one week post-treatment while MPXV-DNA was still detected in the right eye, before reaching undetectable levels four weeks post-treatment. Objective quantification of anterior chamber inflammation through LFP demonstrated gradual decrease that was more pronounced in the affected right eye and coincided with the clinical improvement of the corneal ulcer. CONCLUSIONS: This case of Mpox related corneal ulcer with associated uveitis manifests the feasibility of monitoring the antiviral effect of tecovirimat with virus detection in tear samples and LFP. Our observations indicate that tecovirimat resulted in viral load reduction in both eyes. RT-PCR MPXV detection in tear samples and LFP represent noninvasive tools that could assist with treatment response monitoring.

Computational discovery of tripeptide inhibitors targeting monkeypox virus A42R profilin-like protein.

Monkeypox is an infectious disease caused by the monkeypox virus (MPXV), a member of the Orthopoxvirus genus closely related to smallpox. The structure of the A42R profilin-like protein is the first and only available structure among MPXV proteins. Biochemical studies of A42R were conducted in the 1990s and later work also analyzed the protein's function in viral replication in cells. This study aims to screen tripeptides for their potential inhibition of the A42R profilin-like protein using computational methods, with implications for MPXV therapy. A total of 8000 tripeptides underwent molecular docking simulations, resulting in the identification of 20 compounds exhibiting strong binding affinity to A42R. To validate the docking results, molecular dynamics simulations and free energy perturbation calculations were performed. These analyses revealed two tripeptides with sequences TRP-THR-TRP and TRP-TRP-TRP, which displayed robust binding affinity to A42R. Markedly, electrostatic interactions predominated over van der Waals interactions in the binding process between tripeptides and A42R. Three A42R residues, namely Glu9, Ser12, and Arg38, appear to be pivotal in mediating the interaction between A42R and the tripeptide ligands. Notably, tripeptides containing two or three tryptophan residues demonstrate a pronounced binding affinity, with the tripeptide comprising three tryptophan amino acids showing the highest level of affinity. These findings offer valuable insights for the selection of compounds sharing a similar structure and possessing a high affinity for A42R, potentially capable of inhibiting its enzyme activity. The study highlights a structural advantage and paves the way for the development of targeted therapies against MPXV infections.

Analytical and clinical evaluation of a novel real-time PCR-based detection kit for Mpox virus.

Outbreaks of emerging diseases, like Mpox in 2022, pose unprecedented challenges to global healthcare systems. Although Mpox cases globally decreased since the end of 2022, numbers are still significant in the African Region, European Region, Region of the Americas, and Western Pacific Region. Rapid and efficient detection of infected individuals by precise screening assays is crucial for successful containment. In these assays, analytical and clinical performance must be assessed to ensure high quality. However, clinical studies evaluating Mpox virus (MPXV) detection kits using patient-derived samples are scarce. This study evaluated the analytical and clinical performance of a new diagnostic MPXV real-time PCR detection kit (Sansure Monkeypox Virus Nucleic Acid Diagnostic Kit) using patient-derived samples collected in Germany during the MPXV clade IIb outbreak in 2022. Our experimental approach determined the Limit of Detection (LoD) to less than 200 cp/mL using whole blood samples and samples derived from vesicles or pustules. Furthermore, we tested potentially inhibiting substances and pathogens with homologous nucleic acid sequences or similar clinical presentation and detected no cross-reactivity or interference. Following this, the assay was compared to a CE-marked test in a clinical performance study and achieved a diagnostic sensitivity of 100.00% and diagnostic specificity of 96.97%. In summary, the investigated real-time PCR assay demonstrates high analytical performance and concurs with the competitor device with high specificity and sensitivity.

Human monkeypox virus: A systematic critical review during the pandemic peak.

BACKGROUND: In July 2022, the world health organization (WHO) announced the monkeypox virus (MPXV) as a public health emergency of international concern, due to the unprecedented global transmission of the disease beyond previously endemic countries in Africa. METHODS: For this systematic review, the author searched the "web of science" (WoS) database, which retrieves 138 articles on MPXV, published between 01-04-2022 and 22-09-2022. This period witnessed the maximum cases of infection as confirmed by the WHO. Seventy articles were used for in-depth analysis, after excluding papers not highly relevant to the topic. RESULTS AND CONCLUSIONS: The current review demonstrates different types of MPXV identification analysis, transmission of MPXV, clinical features, immune responses against MPXV, the mutations, and phylogenetic analysis. It also identifies the patients with high-risk complications and determines the other diseases related to MPXV. This paper provides suggestions for the suitable usage of vaccines or antiviral drugs as a procedure to control the outbreak and preventive strategies related to the humans. This research discusses significant implications and recommendations to contribute in reducing the spread of MPXV and presents avenues for upcoming MPXV research.

The Unique Immune Evasion Mechanisms of the Mpox Virus and Their Implication for Developing New Vaccines and Immunotherapies.

Mpox is an infectious and contagious zoonotic disease caused by the mpox virus (MPXV), which belongs to the genus Orthopoxvirus. Since 2022, MPXV has posed a significant threat to global public health. The emergence of thousands of cases across the Western Hemisphere prompted the World Health Organization to declare an emergency. The extensive coevolutionary history of poxviruses with humans has enabled these viruses to develop sophisticated mechanisms to counter the human immune system. Specifically, MPXV employs unique immune evasion strategies against a wide range of immunological elements, presenting a considerable challenge for treatment, especially following the discontinuation of routine smallpox vaccination among the general population. In this review, we start by discussing the entry of the mpox virus and the onset of early infection, followed by an introduction to the mechanisms by which the mpox virus can evade the innate and adaptive immune responses. Two caspase-1 inhibitory proteins and a PKR escape-related protein have been identified as phylogenomic hubs involved in modulating the immune environment during the MPXV infection. With respect to adaptive immunity, mpox viruses exhibit unique and exceptional T-cell inhibition capabilities, thereby comprehensively remodeling the host immune environment. The viral envelope also poses challenges for the neutralizing effects of antibodies and the complement system. The unique immune evasion mechanisms employed by MPXV make novel multi-epitope and nucleic acid-based vaccines highly promising research directions worth investigating. Finally, we briefly discuss the impact of MPXV infection on immunosuppressed patients and the current status of MPXV vaccine development. This review may provide valuable information for the development of new immunological treatments for mpox.

Computational investigations of potential inhibitors of monkeypox virus envelope protein E8 through molecular docking and molecular dynamics simulations.

The World Health Organization (WHO) has declared the monkeypox outbreak a public health emergency, as there is no specific therapeutics for monkeypox virus (MPXV) disease. This study focused on docking various commercial drugs and plant-derived compounds against the E8 envelope protein crucial for MPXV attachment and pathogenesis. The target protein structure was modeled based on the vaccinia virus D8L protein. Notably, maraviroc and punicalagin emerged as potential ligands, with punicalagin exhibiting higher binding affinity (- 9.1 kcal/mol) than maraviroc (- 7.8 kcal/mol). Validation through 100 ns molecular dynamics (MD) simulations demonstrated increased stability of the E8-punicalagin complex, with lower RMSD, RMSF, and Rg compared to maraviroc. Enhanced hydrogen bonding, lower solvent accessibility, and compact motions also attributed to higher binding affinity and stability of the complex. MM-PBSA calculations revealed van der Waals, electrostatic, and non-polar solvation as principal stabilizing energies. The binding energy decomposition per residue favored stable interactions between punicalagin and the protein's active site residues (Arg20, Phe56, Glu228, Tyr232) compared to maraviroc. Overall study suggests that punicalagin can act as a potent inhibitor against MPXV. Further research and experimental investigations are warranted to validate its efficacy and safety.

Role of Direct Sexual Contact in Human Transmission of Monkeypox Virus, Italy.

The 2022 global mpox outbreak was driven by human-to-human transmission, but modes of transmission by sexual relationship versus sexual contact remain unclear. We evaluated sexual transmission of mpox by using monkeypox virus (MPXV) G2R-mRNA as a marker of ongoing viral replication through in vitro experiments. We analyzed clinical samples of 15 MPXV-positive patients in Italy from different biological regions by using the setup method. The presence of MPXV DNA, MPXV G2R-mRNA, or both in all analyzed lesion swab samples, independent of viral load, confirmed a higher infectivity risk from skin lesions. Positivity for MPXV G2R-mRNA in nasopharyngeal swabs was associated with high MPXV load, whereas positive results for MPXV G2R-mRNA were obtained only in the 2 semen samples with the lowest MPXV loads. Our results suggest that close or skin-to-skin contact during sexual intercourse is the main route of sexual transmission and that semen is a minor driver of infection, regardless of MPXV load.

Prevention of sexual transmission of mpox: a systematic review and qualitative evidence synthesis of approaches.

BACKGROUND: The ongoing multi-country mpox outbreak in previously unaffected countries is primarily affecting sexual networks of men who have sex with men. Evidence is needed on the effectiveness of recommended preventive interventions. To inform WHO guidelines, a systematic review and qualitative evidence synthesis were conducted on mpox preventive behavioural interventions to reduce: (i) sexual acquisition; (ii) onward sexual transmission from confirmed/probable cases; and (iii) utility of asymptomatic testing. METHODS: Medline, EMBASE, PubMed, Cochrane and WHO trial databases, grey literature and conferences were searched for English-language primary research published since 1 January 2022. A reviewer team performed screening, data extraction and bias assessment. A qualitative thematic synthesis explored views and experiences of engagement in prevention in individuals at increased risk. RESULTS: There were 16 studies: 1 on contact-tracing, 2 on sexual behaviour, and 13 on asymptomatic testing. Although MPXV was detected in varying proportions of samples (0.17%-6.5%), the testing studies provide insufficient evidence to fully evaluate this strategy. For the qualitative evidence synthesis, four studies evaluated the experiences of most affected communities. Preferences about preventive interventions were shaped by: mpox information; the diversity of sexual practices; accessibility and quality of mpox testing and care; and perceived cost to wellbeing. CONCLUSIONS: Evidence on the effectiveness of interventions to prevent the sexual transmission of mpox remains scarce. Limited qualitative evidence on values and preferences provides insight into factors influencing intervention acceptability. Given global and local inequities in access to vaccines and treatment, further research is needed to establish the effectiveness of additional interventions.

Continued circulation of mpox: an epidemiological and phylogenetic assessment, European Region, 2023 to 2024.

During the summer of 2023, the European Region experienced a limited resurgence of mpox cases following the substantial outbreak in 2022. This increase was characterised by asynchronous and bimodal increases, with countries experiencing peaks at different times. The demographic profile of cases during the resurgence was largely consistent with those reported previously. All available sequences from the European Region belonged to clade IIb. Sustained efforts are crucial to control and eventually eliminate mpox in the European Region.

Inefficient tissue immune response against MPXV in an immunocompromised mpox patient.

The recent outbreak of monkeypox virus (MPXV) was unprecedented in its size and distribution. Those living with uncontrolled HIV and low CD4 T cell counts might develop a fulminant clinical mpox course with increased mortality, secondary infections, and necrotizing lesions. Fatal cases display a high and widespread MPXV tissue burden. The underlying pathomechanisms are not fully understood. We report here the pathological findings of an MPXV-driven abscess in gastrocnemius muscle requiring surgery in an immunocompromised patient with severe mpox. Presence of virus particles and infectivity were confirmed by electron microscopy, expansion microscopy, and virus culture, respectively. MPXV tissue distribution by immunohistochemistry (IHC) showed a necrotic core with infection of different cell types. In contrast, at the lesion rim fibroblasts were mainly infected. Immune cells were almost absent in the necrotic core, but were abundant at the infection rim and predominantly macrophages. Further, we detected high amounts of alternatively activated GPNMB+-macrophages at the lesion border. Of note, macrophages only rarely colocalized with virus-infected cells. Insufficient clearance of infected cells and infection of lesion-associated fibroblasts sustained by the abundance of profibrotic macrophages might lead to the coalescing of lesions and the severe and persistent clinical mpox course observed in immunocompromised patients.

Decoding and Unravelling Mpox, Herpes, and Syphilis Infections: A State of Art Review.

As the world recovers from the COVID-19 pandemic, a resurgence in MPXV cases is causing serious concern. The early clinical similarity of MPXV to common ailments like the flu and cold, coupled with the resemblances of its progressing rash to other infections, underscores the importance of prompt and accurate diagnosis. Among the infections, smallpox is clinically closest to MPXV, and rashes similar to MPXV stages also appear in syphilis and varicella zoster. A comprehensive review of MPXV, herpes, and syphilis was carried out, including structural and morphological features, origins, transmission modes, and computational studies. PubMed literature search on MPXV, using MeSH key terms, yielded 1904 results, with the analysis revealing prominent links to sexually transmitted diseases. More in-depth exploration of MPXV, Herpes Simplex Virus (HSV), and Syphilis revealed further disease interconnections and geographical correlations. These findings emphasize the need for a holistic understanding of these interconnected infectious agents for better control and management.

Functional characterization and structural prediction of hypothetical proteins in monkeypox virus and identification of potential inhibitors.

The excessive activation of the monkeypox virus (MPXV-Congo_8-156) is linked to various skin and respiratory disorders such as rashes, fluid-filled blisters, swollen lymph nodes and encephalitis (inflammation of the brain), highlighting MPXV-Congo_8-156 as a promising target for drug intervention. Despite the effectiveness of Cidofovir, in inhibiting MPXV activity, its limited ability to penetrate the skin and its strong side effects restrict its application. To address this challenge, we screened 500 compounds capable of penetrating the skin and gastrointestinal tract to identify potent MPXV inhibitors. Various characterization schemes and structural models of MPXV-Congo_8-156 were explored with bioinformatics tools like PROTPARAM, SOPMA, SWISS-MODEL and PROCHECK. Using molecular docking in PyRx, we evaluated the binding affinities of these compounds with MPXV-Congo_8-156 and identified the top five candidates ranging from - 9.2 to - 8.8 kcal/mol. ADMET analysis indicated that all five compounds were safer alternatives, showing no AMES toxicity or carcinogenicity in toxicological assessments. Molecular dynamics (MD) simulations, conducted for 100 ns each, confirmed the docking interactions of the top five compounds alongside the control (Cidofovir), validating their potential as MPXV inhibitors. The compounds with PubChem CID numbers 4061636, 4422538, 3583576, 4856107 and 4800629 demonstrated strong support in terms of root-mean-square deviation (RMSD), root-mean-square fluctuation (RMSF), radius of gyration (Rg), solvent-accessible surface area (SASA) value, hydrogen bond analysis, and Molecular Mechanics Poisson-Boltzmann Surface Area (MM-PBSA) analysis. Thus, our investigation identified these five compounds as promising inhibitors of MPXV, offering potential therapeutic avenues. However, further in vivo studies are necessary to validate our findings.

The Re-Emergence of Mpox: Old Illness, Modern Challenges.

The Mpox virus (MPXV) is known to cause zoonotic disease in humans. The virus belongs to the genus Orthopoxvirus, of the family Poxviridae, and was first reported in monkeys in 1959 in Denmark and in humans in 1970 in the Congo. MPXV first appeared in the U.S. in 2003, re-emerged in 2017, and spread globally within a few years. Wild African rodents are thought to be the reservoir of MPXV. The exotic trade of animals and international travel can contribute to the spread of the Mpox virus. A phylogenetic analysis of MPXV revealed two distinct clades (Central African clade and West African clade). The smallpox vaccine shows cross-protection against MPXV infections in humans. Those who have not previously been exposed to Orthopoxvirus infections are more vulnerable to MPXV infections. Clinical manifestations in humans include fever, muscle pain, headache, and vesicle formation on the skin of infected individuals. Pathognomonic lesions include ballooning degenerations with Guarnieri-like inclusions in vesicular epithelial cells. Alterations in viral genome through genetic mutations might favor the re-emergence of a version of MPXV with enhanced virulence. As of November 2023, 92,783 cases and 171 deaths have been reported in 116 countries, representing a global public health concern. Here, we provide insights on the re-emergence of MPXV in humans. This review covers the origin, emergence, re-emergence, transmission, pathology, diagnosis, control measures, and immunomodulation of the virus, as well as clinical manifestations. Concerted efforts of health professionals and scientists are needed to prevent the disease and stop its transmission in vulnerable populations.

The Feasibility of Elimination of Monkeypox Virus in Nigeria: A Systematic Review.

Monkeypox is a zoonotic virus with an increasing incidence in Nigeria, posing significant public health challenges. The virus, related to smallpox, primarily spreads through direct contact with infected animals or humans and has been noted for its potential for wider transmission due to changing ecological and social dynamics. This research aims to evaluate the feasibility of eliminating monkeypox in Nigeria through integrated approaches involving vaccination, public health strategies, and ecological management. Additionally, it seeks to propose a unified public health strategy, incorporating One Health principles, to achieve the elimination of monkeypox in Nigeria. A review of the feasibility of eliminating the monkeypox virus in Nigeria was conducted using databases like Science Direct, Cochrane, PubMed, and Medline. The search was guided by the methodologies and reporting guidelines of the Preferred Reporting Items for Systematic reviews and Meta-Analyses (PRISMA). The initial search yielded 89 publications, but the number of articles that required examination was reduced after applying selected keywords and evaluating abstracts. We deemed 32 articles relevant to the subject after applying inclusion and exclusion criteria. The findings are analyzed, highlighting their limitations and strengths and discussing practical implications, knowledge gaps, and recommendations for future research. This study provides evidence supporting the feasibility of the elimination of the monkeypox virus in Nigeria. Strategic recommendations for a sustainable public health approach to eliminating monkeypox emphasize preventive measures, community engagement, and ecological preservation. This study will provide critical insights into the feasibility of eliminating monkeypox in Nigeria, offering a model that can be adapted for other regions facing similar challenges. The integration of health, ecological, and community-focused strategies is expected to contribute significantly to global efforts to control and potentially eradicate monkeypox. This research could serve as a foundational study for public health authorities in Nigeria and internationally, informing policy and operational decisions to control and eliminate monkeypox as a public health threat. The monkeypox virus reemerges in Nigeria, increasing mortality rates. Awareness programs should be conducted on its danger, transmission mode, and potential human transmission. Public education on the lack of treatment and vaccines is crucial. Meat inspection laws should be enforced to ensure safe animal consumption.

No evidence of mpox virus circulation in putative animal reservoirs in Gabon wildlife.

OBJECTIVES: Mpox is a neglected viral endemic tropical disease in Central and Western African countries transmitted to humans by an animal. However, the natural reservoir of the virus remains elusive. In this study, we looked for potential reservoirs of the mpox virus (MPXV) in Gabonese wildlife to prevent future outbreaks and enrich the literature with additional data on animal reservoirs. METHODS: DNA was extracted from the livers and spleens from 2549 animals (bats [859], bushmeats [356], rodents [1309], and shrews [25]) collected between 2012 and 2021. DNA was analyzed by real-time and conventional polymerase chain reaction, targeting the 14 kD protein and the rpo subunit RNA polymerase of orthopoxviruses. RESULTS: No MPXV DNA was detected despite the presence of potential host reservoirs such as Critcetomys, Crocidura, Praomys, and Atherurus africanus. This absence could be due to (i) the low number of animals collected for some species, (ii) the acute nature of mpox infection but also (iii) the lack of the potential reservoir Funisciurus anerythrus among collected animals, and (iv) the fact that the samplings are not included in the probable ecological niche of MPXV. CONCLUSION: Longitudinal studies including potential ecological niches of F. anerythrus and MPXV in Gabon may be useful to get more information on MPXV circulation.

Clinically inapparent mpox virus (MPXV) infections among clients of three anonymous Community Based Voluntary Counselling and Testing centres in Berlin, Germany, 2022-2023.

INTRODUCTION: Since the mpox outbreak in 2022, it was unclear if and how often infections with mpox virus (MPXV) were clinically inapparent, i.e. not presenting to clinical care with mpox symptoms. Moreover, it was hypothesized that MPXV circulated in the affected communities before the outbreak was officially detected. METHODS: We retrospectively tested rectal and urethral swabs, and pooled samples for presence of MPXV. Samples were obtained from routine STI testing of three anonymous Community Based Voluntary Counselling and Testing (CBVCT) centres in Berlin, in 2022 and 2023. Testing results were linked to anonymously provided behavioural data. RESULTS: Overall, 9,053 samples from 6,600 client visits were included. Clinically inapparent MPXV infections were detectable in 1.1% of the samples. We did not find MPXV infections in the month before the first cases appeared in Berlin or between October 2022 and January 2023 when case numbers were low in Germany. However, during the outbreak period in 2022, we found clinically inapparent MPXV infections among 2.2% of the clients and during summer/autumn 2023 among 0.3%. The number of condomless anal/vaginal intercourse partners within the previous 6 months and PrEP use were identified as predictors of clinically inapparent MPXV infection. CONCLUSION: Clinically inapparent MPXV infections occurred during the mpox outbreak in Berlin in 2022 and post-outbreak in summer/autumn 2023. Unrecognized MPXV circulation in Berlin before the recognition of the outbreak in May 2022 appears unlikely. However, low-level sustained circulation of clinically inapparent MPXV infections need to be acknowledged in mpox prevention strategies.

Using wastewater surveillance for mpox as a complement to traditional case-based reporting - Chicago, March-June 2023.

The Chicago Department of Public Health tested wastewater samples for the presence of Monkeypox Virus (MPXV) from March 13 through June 26, 2023. There were persistent detections prior to reported cases. This indicated the baseline levels of MPXV prevalence might warrant routine monitoring. Detections in areas without corresponding reported clinical cases might highlight areas where cases are being under-reported by traditional surveillance.

Reemergence of Clade IIb-Associated Mpox, Germany, July-December 2023.

In July 2023, clade IIb-associated mpox reemerged in Germany at low levels, mainly affecting men who have sex with men. We report a representative case and phylogeny of available genome sequences. Our findings underscore the need for standardized surveillance and indication-based vaccination to limit transmission and help prevent endemicity.

Zoonotic and Zooanthroponotic Potential of Monkeypox.

The current multicounty outbreak of monkeypox virus (MPXV) posed an emerging and continued challenge to already strained public healthcare sector, around the globe. Since its first identification, monkeypox disease (mpox) remained enzootic in Central and West African countries where reports of human cases are sporadically described. Recent trends in mpox spread outside the Africa have highlighted increased incidence of spillover of the MPXV from animal to humans. While nature of established animal reservoirs remained undefined, several small mammals including rodents, carnivores, lagomorphs, insectivores, non-human primates, domestic/farm animals, and several species of wildlife are proposed to be carrier of the MPXV infection. There are established records of animal-to-human (zoonotic) spread of MPXV through close interaction of humans with animals by eating bushmeat, contracting bodily fluids or trading possibly infected animals. In contrast, there are reports and increasing possibilities of human-to-animal (zooanthroponotic) spread of the MPXV through petting and close interaction with pet owners and animal care workers. We describe here the rationales and molecular factors which predispose the spread of MPXV not only amongst humans but also from animals to humans. A range of continuing opportunities for the spread and evolution of MPXV are discussed to consider risks beyond the currently identified groups. With the possibility of MPXV establishing itself in animal reservoirs, continued and broad surveillance, investigation into unconventional transmissions, and exploration of spillover events are warranted.