Harnessing SuperNatural database to identify VP35 inhibitors as anti-Ebola drug candidates: A multistage In silico study.

Analyzing The Ebola ViruAnalyzing The Impact That The Ebola Virus Disease Has on Children

The recent outbreak of the Ebola virus (EBOV) has marked it as one of the most severe health threats globally. Among various anti-EBOV inhibitors studied, galidesivir (BCX4430) has shown remarkable efficacy. This study aims to identify novel potential anti-EBOV drugs among galidesivir analogs, focusing on the Zaire ebolavirus (Z-EBOV), which exhibits a mortality rate of 90%. We subjected 200 candidate compounds to molecular docking calculations, followed by an evaluation of the bioactivity of the top 25 compounds using the OSIRIS Property Explorer. Initial 50 ns molecular dynamics (MD) simulations were then performed. According to our findings, only six compounds exhibited positive drug scores. We further performed molecular mechanics Poisson-Boltzmann surface area (MM/PBSA) calculations of binding energy over 50 ns, selecting the two top-performing compounds for extended 150 ns MD simulations. CID 117698807 and CID 117712809 showed higher binding stability compared to galidesivir, with ΔG binding values of −36.7 and −53.4 kcal/mol, respectively. Both compounds demonstrated high stability within the Z-EBOV-V24 active site over the 150 ns MD simulations. Hence, our study proposes CID 117698807 and CID 117712809 as potential anti-Z-EBOV-V24 drug candidates, warranting further investigation. Communicated by Ramaswamy H. Sarma

Peter L Collins & Alexander Bukreyev† †Author for correspondence Laboratory of Infectious Disease, National Institute of Allergy & Infectious Diseases, National Institutes of Health, 50 South Drive, Room 6505, Bethesda, MA 20892-8007, USA Tel.: +1 301 594 1854; Fax: +1 301 496 8312; ab176v@nih.gov ‘The development of vaccines against MARV and EBOV ... is a high priority because of their high virulence, ability to spread from person to person and the possibility of their use in bioterrorism.’

Providing nursing care to Ebola patients on the national stage: The National Institutes of Health experience

Are adaptive randomised trials or non-randomised studies the best way to address the Ebola outbreak in west Africa?

Objective The aim of this study is to analyze the clinical features of patients with Ebola virus (EBOV) infection at admission and to provide information for clinicians to manage patients with suspected Ebola virus disease (EVD). Methods Sixty-two suspected EVD cases visited China-Sierra leone Friendship Hospital which was set up by the first medical team from the Chinese People′s Liberation Army in Africa were enrolled in this study. Demographic information, medical history and symptoms were collected for all the patients at admission. Reverse transcription polymerase chain reaction (RT-PCR) was used to confirm or exclude the diagnosis. Data were compared between patients with or without EBOV infection by t test for ages, days from symptoms onset to admission, and χ2 test for proportion of symptoms. Results Of 62 patients, 35 were male and 27 were female, average age was (27.3±15.4) years. Forty cases were diagnosed with EVD, and 22 were excluded. The incubation period for EVD patients was estimated to be 2 to 24 days with an average of (8.8±5.7) days. EBOV positive patients showed higher proportions of fever, nausea/vomiting, diarrhea, fatigue/weakness, loss of appetite and sore throat than those of EBOV negative patients (all P<0.05). The proportions of eye pain, hiccup and insomnia were low in EBOV positive patients, while none was in EBOV negative patients. Only one patient had bloody stool at admission. An average of (8±5) symptoms were noted in EVD patients at admission compared with (4±3) symptoms in non-EVD patients (t=8.46, P=0.001). The number of symptoms at admission was optimal in diagnosing EVD with cut-off value of 6. The sensitivity, specificity, positive predictive value and negative predictive value were 50.0%, 86.4%, 87.0% and 48.7%, respectively. Thirty-six out of 62 patients had a history of contact with confirmed or suspected EVD cases, including 22(55.0%) in EBOV positive patients and 14(63.6%) in EBOV negative patients (χ2=0.44, P=0.596). Among 36 cases with definite EVD contact history, 19(86.4%) presented with fever and 20(90.9%) presented with fatigue in EBOV positive patients, while 6(42.9%) presented with fever and 5(35.7%) presented with fatigue in EBOV negative patients (both P<0.05). Conclusions Among suspected EVD patients, those who are diagnosed with EVD by RT-PCR have more symptoms than those with negative results, including fever, nausea/vomiting, diarrhea, fatigue/weakness, loss of appetite and sore throat, etc. Though these symptoms are not specific, they provide clues for diagnose EVD in the very early stage for clinicians. Key words: Hemorrhagic fever, ebola; Holding center; Clinical features

Ebola and the Ophthalmologist

Kenema Government Hospital (KGH) has developed an advanced clinical and laboratory research capacity to manage the threat of Lassa fever, a viral hemorrhagic fever (VHF). The 2013-2016 Ebola virus (EBOV) disease (EVD) outbreak is the first to have occurred in an area close to a facility with established clinical and laboratory capacity for study of VHFs. Because of its proximity to the epicenter of the EVD outbreak, which began in Guinea in March 2014, the KGH Lassa fever Team mobilized to establish EBOV surveillance and diagnostic capabilities. Augustine Goba, director of the KGH Lassa laboratory, diagnosed the first documented case of EVD in Sierra Leone, on 25 May 2014. Thereafter, KGH received and cared for numbers of patients with EVD that quickly overwhelmed the capacity for safe management. Numerous healthcare workers contracted and lost their lives to EVD. The vast majority of subsequent EVD cases in West Africa can be traced back to a single transmission chain that includes this first diagnosed case. Responding to the challenges of confronting 2 hemorrhagic fever viruses will require continued investments in the development of countermeasures (vaccines, therapeutic agents, and diagnostic assays), infrastructure, and human resources.

Emerging Pathogens: Challenges and Successes of Molecular Diagnostics

Ebola virus (EBOV) is the causative agent of severe hemorrhagic fever in primates, with human case fatality rates up to 90%. Today, there is neither a licensed vaccine nor a treatment available for Ebola hemorrhagic fever (EHF). Single monoclonal antibodies (MAbs) specific for Zaire ebolavirus (ZEBOV) have been successfully used in passive immunization experiments in rodent models, but have failed to protect nonhuman primates from lethal disease. In this study, we used two clones of human-mouse chimeric MAbs (ch133 and ch226) with strong neutralizing activity against ZEBOV and evaluated their protective potential in a rhesus macaque model of EHF. Reduced viral loads and partial protection were observed in animals given MAbs ch133 and ch226 combined intravenously at 24 hours before and 24 and 72 hours after challenge. MAbs circulated in the blood of a surviving animal until virus-induced IgG responses were detected. In contrast, serum MAb concentrations decreased to undetectable levels at terminal stages of disease in animals that succumbed to infection, indicating substantial consumption of these antibodies due to virus replication. Accordingly, the rapid decrease of serum MAbs was clearly associated with increased viremia in non-survivors. Our results indicate that EBOV neutralizing antibodies, particularly in combination with other therapeutic strategies, might be beneficial in reducing viral loads and prolonging disease progression during EHF.

During an Ebola virus disease (EVD) outbreak, health workers (HWs) are at high risk of EVD infection. Health workers infected with EVD are a major risk factor for the failure to control EVD outbreaks. The tenth outbreak is the largest recorded in the DRC, but unfortunately few studies have described the occurrence, case-fatality ratio (CFR) among HWs and their perceptions. The aim of this study was to describe: the occurrence of EVD; related mortality; and perceptions of EVD transmission among HWs. A mixed methods study was conducted in north-eastern DRC in August 2019. A review of EVD surveillance data and in depth interviews with 16 front line HWs were carried out. Interviews were audio recorded and transcribed verbatim in Swahili before being translated into French and subsequently into English. Quantitative data were imported from Microsoft Excel to Stata 14.0 for analysis. Mean and standard deviation (SD) were used to summarize numeric variables while frequency was used for categorical data. All descriptive statistics are reported with their 95% confidence interval. From 12 July 2018 to 17 August 2019, some 151 confirmed and three probable health worker cases were recorded in the viral haemorrhagic fever (VHF) surveillance dataset for the provinces of North-Kivu and Ituri. Heath workers accounted for 5.5% [4.6-6.4]. More than half of the HWs infected were male, with a male to female ratio of 1.3. The mean age of HWs infected was 33.9 years (sd: 10.3). Nosocomial transmission was the most common route of transmission among HWs (94.1%). For more than a quarter of the HWs the contact case was unknown. From a total of 154 infected HWs, some 29 died (CFR: 18.8% [13.0-25.9]). The factors mentioned by HWs as being associated with EVD transmission among them were: ignorance that patient was infected; unavailability of protective equipment; non-compliance of hygienic measures like regular hand washing; violation of EVD infection prevention and control (IPC) standards; and their resistance thereto. The occurrence of EVD cases among HWs is a challenge for the control of the current outbreak in the DRC. It is important to implement strategies to reduce transmission in this population group.

Infection with Marburg and Ebola viruses cause haemorrhagic fevers that are characterized by organ malfunction, bleeding complications, and high mortality. The viruses are members of the family Filoviridae, a group of membrane-enveloped filamentous RNA viruses. Five distinct species of the genus Ebolavirus have been reported; the genus Marburgvirus contains only one species. Both Marburg and Ebola virus diseases are zoonotic infections whose primary hosts are thought to be bats. The initial human infection is acquired from wildlife and subsequent person-to-person spread propagates the outbreak until it is brought under control. Ebola and Marburg viruses are classified as hazard or risk group 4 pathogens because of the very high case fatality rates observed for Ebola and Marburg virus diseases, the frequency of person-to-person transmission and community spread, and the lack of an approved vaccine or antiviral therapy. This mandates that infectious materials are handled and studied in maximum containment laboratory facilities. Epidemics have occurred sporadically since the discovery of Marburg in 1967 and Ebola virus in 1976. While some of these outbreaks have been relatively large, infecting a few hundreds of individuals, they have generally occurred in rural settings and have been controlled relatively easily. However, the 2013–2016 epidemic of Ebola virus disease in West Africa was different, representing the first emergence of the Zaire species of Ebola in a high-density urban location. Consequently, this has been the largest recorded filovirus outbreak in both the number of people infected and the range of geographical spread. Many of the reported and confirmed cases were among people living in high-density and impoverished urban environments. The chapter summarizes the most up-to-date taxonomic status of the family Filoviridae. It focuses on Marburg and Ebola viruses in a historical context, culminating in the 2013–2016 outbreak of Ebola virus in West Africa. Virus biology of the most well-studied member is described, with details of the viral genome and the protein machinery necessary to propagate viruses at the molecular and cellular level. This information is used to build a wider-scale virus–host perspective with detail on the pathology and pathogenesis of Ebola virus disease. The consequences of cell infection are examined, together with our current understanding of the immune response to Ebola virus, leading to a broader description of the clinical features of disease. The chapter closes by drawing information together in a section on diagnosis, ecology, prevention, and control.

Ebola virus (EBOV) is a human pathogen with the ability to cause hemorrhagic fever and bleeding diathesis in hosts. The life cycle of EBOV depends on its nucleocapsid. The Ebola nucleocapsid consists of a helical assembly of nucleoproteins (NPs) encapsidating single-stranded viral RNA (ssRNA). Knowledge of the molecular determinants of Ebola nucleocapsid stability is essential for the development of therapeutics against EBOV. However, large degrees of freedom associated with the Ebola nucleocapsid helical assembly pose a computational challenge, thereby limiting the previous simulation studies to the level of monomers. In the present work, we have performed all atom molecular dynamics (MD) simulations of the helical assembly of EBOV nucleoproteins in the absence and presence of ssRNA. We found that ssRNA is essential for maintaining structural integrity of the nucleocapsid. Other molecular determinants observed to stabilize the nucleocapsid include NP-RNA and NP-NP interactions and ion distributions. Additionally, the structural and dynamical behavior of the nucleocapsid monomer depends on its position in the helical assembly. NP monomers present on the longitudinal edges of the helical tube are more exposed, flexible, and have weaker NP-NP interactions than those residing in the center. This work provides key structural features stabilizing the nucleocapsid that may serve as therapeutic targets.

Ebola virus (EBOV), one of the deadliest diseases, is responsible for infecting individuals with hemorrhagic fever syndrome, which remains an ongoing worldwide health concern. The extremely deadly nature and virulence of EBOV illness illuminate the imperative need to evolve effective curative agents. Viral protien (VP35) acts as an Achilles heel for EBOV reproduction and also interacts with numerous human proteins, which leads to impairing the immune system. Herein, the DrugBank database, containing >14000 investigational and approved drugs, is mined to hunt prospective inhibitors toward VP35 utilizing various computational approaches. Docking technique performance is initially validated to predict the VP35‐inhibitor binding pose upon the accessible experimental data. Molecular dynamics simulations (MDS) are then conducted in triplicate on the top potent drug candidates, followed by binding energy (ΔGbinding) estimations using molecular mechanics/generalized Born surface area (MM/GBSA) approach. Upon MM/GBSA//250 ns MDS, DB14875 and DB07800 revealed better binding energy against VP35 than 1D9, reference inhibitor, with ΔGbinding values of −36.6, −35.6, and −29.3 kcal mol−1, respectively. Post‐MD analyses demonstrate great stability for the identified drug candidates complexed with VP35 over 250 ns MDS. Ultimately, the density functional theory computations are executed, and their outcomes elucidate favorable molecular reactivity of the identified drug candidates. Conclusively, these findings suggest promising inhibitors for VP35, warranting further experimental assays.

Recently, there have been substantial developments in the understanding of Ebola hemorrhagic fever pathogenesis, but there are still major gaps. These infections occur in underdeveloped areas of the world, and much of our knowledge of naturally occurring disease is derived from sporadic outbreaks that occurred decades in the past. Recently conducted laboratory animal studies have provided insight into Ebola pathogenesis and may help guide clinical investigations of disease using contemporary methodologies that were not available previously. A better understanding of the relevant host and viral factors that influence clinical and virologic outcome will be critical to our ability to combat this aggressive pathogen. This article reviews the most relevant information relating to the postulated pathogenesis of this disease, focusing on the role of the endothelium in contributing to the coagulation disorders that characterize Ebola hemorrhagic fever in primates. Some of the remaining and key unanswered questions relating to the role of the vascular system in the pathogenesis of this disease, that need to be addressed in further research, are highlighted.