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Background: Individuals may exhibit altered foot pronation during gait when fatigue sets in. Therefore, a more evidence-based understanding of these fatigue-induced changes may be helpful for future gait analysis and return-to-play tests since fatigue can provide new insights that might explain a person’s complaints. Methods: A total of 25 healthy individuals (12♂, 13♀; 24.3 ± 2.7 years; 174.9 ± 9.09 cm; 70 ± 14.2 kg; BMI: 22.7 ± 2.8) participated in this controlled non-randomized study of unilateral fatigue of the right foot’s stabilizing muscles with regard to the pronation of the foot, measured by navicular drop (ND) in static (statND; standing) and dynamic (dynND; walking) states. The left foot served as the control. Surface electromyography was used to verify fatigue. Results: While the statND did not change, the dynND increased significantly by 1.44 ± 2.1 mm (=22.3%) after the foot-stabilizing muscles experienced fatigue. No correlation was found between the statND and dynND. Conclusions: Muscular fatigue can affect foot pronation. The dynND appears to be more representative of the loads in everyday life, whereby most studies use the statND.

Summary: Besides neutralizing antibodies, which are considered an important measure for vaccine immunogenicity, Fc-mediated antibody functions can contribute to antibody-mediated protection. They are strongly influenced by structural antibody properties such as subclass and Fc glycan composition. We here applied a systems serology approach to dissect humoral immune responses induced by MVA-MERS-S, an MVA-vectored vaccine against the Middle East respiratory syndrome coronavirus (MERS-CoV). Building on preceding studies reporting the safety and immunogenicity of MVA-MERS-S, our study highlights the potential of a late boost, administered one year after prime, to enhance both neutralizing and Fc-mediated antibody functionality compared to the primary vaccination series. Distinct characteristics were observed for antibodies specific to the MERS-CoV spike protein S1 and S2 subunits, regarding subclass and glycan compositions as well as Fc functionality. These findings highlight the benefit of a late homologous booster vaccination with MVA-MERS-S and may be of interest for the design of future coronavirus vaccines.

Abstract In response to the COVID-19 pandemic, multiple vaccines were developed using platforms such as viral vectors and mRNA technology. Here, we report humoral and cellular immunogenicity data from human phase 1 clinical trials investigating two recombinant Modified Vaccinia virus Ankara vaccine candidates, MVA-SARS-2-S and MVA-SARS-2-ST, encoding the native and the prefusion-stabilized SARS-CoV-2 spike protein, respectively. MVA-SARS-2-ST was more immunogenic than MVA-SARS-2-S, but both were less immunogenic compared to licensed mRNA- and ChAd-based vaccines in SARS-CoV-2 naïve individuals. In heterologous vaccination, previous MVA-SARS-2-S vaccination enhanced T cell functionality and MVA-SARS-2-ST boosted the frequency of T cells and S1-specific IgG levels when used as a third vaccination. While the vaccine candidate containing the prefusion-stabilized spike elicited predominantly S1-specific responses, immunity to the candidate with the native spike was skewed towards S2-specific responses. These data demonstrate how the spike antigen conformation, using the same viral vector, directly affects vaccine immunogenicity in humans.

Objective: Biomechanical Machine Learning (ML) models, particularly deep-learning models, demonstrate the best performance when trained using extensive datasets. However, biomechanical data are frequently limited due to diverse challenges. Effective methods for augmenting data in developing ML models, specifically in the human posture domain, are scarce. Therefore, this study explored the feasibility of leveraging generative artificial intelligence (AI) to produce realistic synthetic posture data by utilizing three-dimensional posture data.Methods: Data were collected from 338 subjects through surface topography. A Variational Autoencoder (VAE) architecture was employed to generate and evaluate synthetic posture data, examining its distinguishability from real data by domain experts, ML classifiers, and Statistical Parametric Mapping (SPM). The benefits of incorporating augmented posture data into the learning process were exemplified by a deep autoencoder (AE) for automated feature representation.Results: Our findings highlight the challenge of differentiating synthetic data from real data for both experts and ML classifiers, underscoring the quality of synthetic data. This observation was also confirmed by SPM. By integrating synthetic data into AE training, the reconstruction error can be reduced compared to using only real data samples. Moreover, this study demonstrates the potential for reduced latent dimensions, while maintaining a reconstruction accuracy comparable to AEs trained exclusively on real data samples.Conclusion: This study emphasizes the prospects of harnessing generative AI to enhance ML tasks in the biomechanics domain.

Competitive climbers engage in highly structured training regimens to achieve peak performance levels, with efficient time management as a critical aspect. Neuromuscular electrical stimulation (NMES) training can close the gap between time-efficient conditioning training and achieving optimal prerequisites for peak climbing-specific performances. Therefore, we examined potential neuromuscular adaptations resulting from the NMFES intervention by analyzing the efficacy of twice-weekly NMES-supported fingerboard (hang board) training compared with thrice-weekly conventional fingerboard training over 7 training weeks in enhancing climbing-specific endurance among intermediate to advanced climbers. Participants were randomly divided into the NMES and control groups. Eighteen participants completed the study (14 male, 4 female; mean age: 25.7 ± 5.3 years; mean climbing experience: 6.4 ± 3.4 years). Endurance was assessed by measuring the maximal time athletes could support their body weight (hanging to exhaustion) on a 20 mm-deep ledge at three intervals: pre-, in-between- (after 4 weeks of training), and post-training (after 7 weeks of training). The findings revealed that despite the lower training volume in the NMES group, no significant differences were observed between the NMES and control groups in climbing-specific endurance. Both groups exhibited notable improvements in endurance, particularly after the in-between test. Consequently, a twice-weekly NMES-supported fingerboard training regimen demonstrated non-inferiority to a thrice-weekly conventional training routine. Incorporating NMES into fingerboard workouts could offer time-saving benefits.

IntroductionThe use of single supplements as ergogenic aids to enhance performance in strength-oriented sports is widespread among athletes (74%). The aim of this study was to increase knowledge about the combined effects of caffeine and L-arginine dietary supplements on performance.MethodsIn this double-blind, randomized and counterbalanced crossover study, 29 participants (age: 23.2 ± 3.6 yr.; height: 181.0 ± 7.6 cm; weight: 77.0 ± 8.8 kg) each underwent six trials. In each trial performance tests were conducted to examine the effects of the supplement combinations on maximum (NmMax) and averaged torque (NmM), maximum (JMax) and averaged work (JM), the blockwise mean values of torque and work, and rate of perceived exertion (RPE) during an isokinetic leg extension task (90°·s−1) with the right leg for two sets of 40 repetitions and a set rest of 3 min on a dynamometer. The first and second trials were used to familiarize the participants with the movements in the dynamometer and no supplements were taken. After this 2-week pre-test trial, the supplement combinations of placebo/placebo, caffeine/placebo (5 mg·kg−1), L-arginine/placebo (0.15 g·kg−1), and caffeine/L-arginine (5 mg·kg−1 + 0.15 g·kg−1) were ingested.ResultsThe main finding of this study is the absence of an ergogenic effect of the combined supplements caffeine and L-arginine during voluntary maximal isokinetic leg extensions, although an increase of 3.5% was noted for Nmmax compared to the placebo trial. However, the administration of caffeine was able to increase the NmMax of the quadriceps femoris muscle about 5.1% (p = 0.043). In addition, caffeine (4.2%, p = 0.026) and also L-arginine (4.2%, p = 0.040) significantly increased NmM over a complete set. No single or combined supplement had an effect on muscle fatigue looking at the blockwise mean values of torque and work or RPE (all p > 0.05).ConclusionThe combination of caffeine and L-arginine was not superior to the isolated intake of both supplements in a strength-based exercise and a synergistic effect was absent.

(1) Background: Non-surgical endodontic treatment has been shown to be clinically successful; however, clinical long-term data are scarce. This practice-based retrospective clinical investigation evaluated endodontic outcomes over 40 years and identified relevant clinical co-factors. (2) Methods: Two experienced dental practitioners in two different private dental practices treated 174 patients with 245 teeth from 1969 to 1993. After root canal obturation, either a new direct restoration (amalgam, resin composite, or glass-ionomer cement) or the re-cementation of a pre-existing prosthetic restoration or renewal of prosthetic restoration followed. Metal posts (operator A) or metal screws (operator B) were inserted when coronal substance loss was significant. The primary outcome (i.e., tooth survival) was achieved when the endodontically treated tooth was, in situ, painless and had full function at the end of the observation period. A secondary outcome, the impact of different prognostic factors on survival rate, was evaluated. (3) Results: The overall mean survival was 56.1% of all treated teeth after 40 years of clinical service, resulting in an annual failure rate of 1.1%. Most investigated clinical co-factors (jaw, tooth position, intracanal dressings, post/screw placement, and gender) showed no significant influence on survival. (4) Conclusions: Even with materials and techniques from the 1970s and 1980s, successful root canal treatment was achievable. Except for post-endodontic restorations, most of the evaluated factors had no significant influence on the clinical long-term survival of root canal-treated teeth.

(1) Background: Occupational fatigue is a primary factor leading to work-related musculoskeletal disorders (WRMSDs). Kinematic and kinetic experimental studies have been able to identify indicators of WRMSD, but research addressing real-world workplace scenarios is lacking. Hence, the authors of this study aimed to assess the influence of physical strain on the Borg CR-10 body map, ergonomic risk scores, and foot pressure in a real-world setting. (2) Methods: Twenty-four participants (seventeen men and seven women) were included in this field study. Inertial measurement units (IMUs) (n = 24) and in-shoe plantar pressure measurements (n = 18) captured the workload of production and office workers at the beginning of their work shift and three hours later, working without any break. In addition to the two 12 min motion capture processes, a Borg CR-10 body map and fatigue visual analog scale (VAS) were applied twice. Kinematic and kinetic data were processed using MATLAB and SPSS software, resulting in scores representing the relative distribution of the Rapid Upper Limb Assessment (RULA) and Computer-Assisted Recording and Long-Term Analysis of Musculoskeletal Load (CUELA), and in-shoe plantar pressure. (3) Results: Significant differences were observed between the two measurement times of physical exertion and fatigue, but not for ergonomic risk scores. Contrary to the hypothesis of the authors, there were no significant differences between the in-shoe plantar pressures. Significant differences were observed between the dominant and non-dominant sides for all kinetic variables. (4) Conclusions: The posture scores of RULA and CUELA and in-shoe plantar pressure side differences were a valuable basis for adapting one-sided requirements in the work process of the workers. Traditional observational methods must be adapted more sensitively to detect kinematic deviations at work. The results of this field study enhance our knowledge about the use and benefits of sensors for ergonomic risk assessments and interventions.

In a clinical trial, Fathi et al. show that a booster vaccination with a vector vaccine candidate against the highly pathogenic Middle East Respiratory Syndrome coronavirus is safe and strongly improves the immunity generated by primary immunization.

Marburg virus causes severe hemorrhagic fever in humans. Here, the authors determine the structure of Marburg virus nucleoprotein–RNA complex by cryo-electron microscopy and provide mechanistic insight into the helical assembly of the nucleocapsid.

The successful development of effective viral vaccines depends on well-known correlates of protection, high immunogenicity, acceptable safety criteria, low reactogenicity, and well-designed immune monitoring and serology. Virus-neutralizing antibodies are often a good correlate of protective immunity, and their serum concentration is a key parameter during the pre-clinical and clinical testing of vaccine candidates. Viruses are inherently infectious and potentially harmful, but we and others developed replication-defective SARS-CoV-2 virus-like-particles (VLPs) as surrogates for infection to quantitate neutralizing antibodies with appropriate target cells using a split enzyme-based approach. Here, we show that SARS-CoV-2 and Epstein–Barr virus (EBV)-derived VLPs associate and fuse with extracellular vesicles in a highly specific manner, mediated by the respective viral fusion proteins and their corresponding host receptors. We highlight the capacity of virus-neutralizing antibodies to interfere with this interaction and demonstrate a potent application using this technology. To overcome the common limitations of most virus neutralization tests, we developed a quick in vitro diagnostic assay based on the fusion of SARS-CoV-2 VLPs with susceptible vesicles to quantitate neutralizing antibodies without the need for infectious viruses or living cells. We validated this method by testing a set of COVID-19 patient serum samples, correlated the results with those of a conventional test, and found good sensitivity and specificity. Furthermore, we demonstrate that this serological assay can be adapted to a human herpesvirus, EBV, and possibly other enveloped viruses.

The SARS-CoV-2 spike (S) glycoprotein is synthesized as a large precursor protein and must be activated by proteolytic cleavage into S1 and S2. A recombinant modified vaccinia virus Ankara (MVA) expressing native, full-length S protein (MVA-SARS-2-S) is currently under investigation as a candidate vaccine in phase I clinical studies. Initial results from immunogenicity monitoring revealed induction of S-specific antibodies binding to S2, but low-level antibody responses to the S1 domain. Follow-up investigations of native S antigen synthesis in MVA-SARS-2-S–infected cells revealed limited levels of S1 protein on the cell surface. In contrast, we found superior S1 cell surface presentation upon infection with a recombinant MVA expressing a stabilized version of SARS-CoV-2 S protein with an inactivated S1/S2 cleavage site and K986P and V987P mutations (MVA-SARS-2-ST). When comparing immunogenicity of MVA vector vaccines, mice vaccinated with MVA-SARS-2-ST mounted substantial levels of broadly reactive anti-S antibodies that effectively neutralized different SARS-CoV-2 variants. Importantly, intramuscular MVA-SARS-2-ST immunization of hamsters and mice resulted in potent immune responses upon challenge infection and protected from disease and severe lung pathology. Our results suggest that MVA-SARS-2-ST represents an improved clinical candidate vaccine and that the presence of plasma membrane–bound S1 is highly beneficial to induce protective antibody levels.

Here, Shaban et al. show that coronaviruses modulate ER stress and the unfolded protein response. The ER stress inducer thapsigargin exerts potent antiviral effects, partially reverses the virus-induced translational shut-down, reprograms the host proteome and suppresses autophagic flux, thereby inhibiting coronavirus replication at multiple levels.

The emergence of Marburg virus (MARV) in Guinea and Ghana triggered the assembly of the MARV vaccine “MARVAC” consortium representing leaders in the field of vaccine research and development aiming to facilitate a rapid response to this infectious disease threat. Here, we discuss current progress, challenges, and future directions for MARV vaccines.

This pilot study aimed to investigate the use of sensorimotor insoles in pain reduction, different orthopedic indications, and the wearing duration effects on the development of pain. Three hundred and forty patients were asked about their pain perception using a visual analog scale (VAS) in a pre–post analysis. Three main intervention durations were defined: VAS_post: up to 3 months, 3 to 6 months, and more than 6 months. The results show significant differences for the within-subject factor “time of measurement”, as well as for the between-subject factor indication (p < 0.001) and worn duration (p < 0.001). No interaction was found between indication and time of measurements (model A) or between worn duration and time of measurements (model B). The results of this pilot study must be cautiously and critically interpreted, but may support the hypothesis that sensorimotor insoles could be a helpful tool for subjective pain reduction. The missing control group and the lack of confounding variables such as methodological weaknesses, natural healing processes, and complementary therapies must be taken into account. Based on these experiences and findings, a RCT and systematic review will follow.

The COVID-19 pandemic caused significant human health and economic consequences. Due to the ability of SARS-CoV-2 to spread rapidly and to cause severe disease and mortality in certain population groups, vaccines are essential for controlling the pandemic in the future. Several licensed vaccines have shown improved protection against SARS-CoV-2 after extended-interval prime-boost immunizations in humans. Therefore, in this study, we aimed to compare the immunogenicity of our two Modified Vaccinia virus Ankara (MVA) based COVID-19 candidate vaccines MVA-SARS-2-S and MVA-SARS-2-ST after short- and long-interval prime-boost immunization schedules in mice. We immunized BALB/c mice using 21-day (short-interval) or 56-day (long-interval) prime-boost vaccination protocols and analyzed spike (S)-specific CD8 T cell immunity and humoral immunity. The two schedules induced robust CD8 T cell responses with no significant differences in their magnitude. Furthermore, both candidate vaccines induced comparable levels of total S, and S2-specific IgG binding antibodies. However, MVA-SARS-2-ST consistently elicited higher amounts of S1-, S receptor binding domain (RBD), and SARS-CoV-2 neutralizing antibodies in both vaccination protocols. Overall, we found very comparable immune responses following short- or long-interval immunization. Thus, our results suggest that the chosen time intervals may not be suitable to observe potential differences in antigen-specific immunity when testing different prime-boost intervals with our candidate vaccines in the mouse model. Despite this, our data clearly showed that MVA-SARS-2-ST induced superior humoral immune responses relative to MVA-SARS-2-S after both immunization schedules.

Filovirus-infected cells are characterized by typical cytoplasmic inclusion bodies (IBs) located in the perinuclear region. The formation of these IBs is induced mainly by the accumulation of the filoviral nucleoprotein NP, which recruits the other nucleocapsid proteins, the polymerase co-factor VP35, the polymerase L, the transcription factor VP30 and VP24 via direct or indirect protein-protein interactions. Replication of the negative-strand RNA genomes by the viral polymerase L and VP35 occurs in the IBs, resulting in the synthesis of positive-strand genomes, which are encapsidated by NP, thus forming ribonucleoprotein complexes (antigenomic RNPs). These newly formed antigenomic RNPs in turn serve as templates for the synthesis of negative-strand RNA genomes that are also encapsidated by NP (genomic RNPs). Still in the IBs, genomic RNPs mature into tightly packed transport-competent nucleocapsids (NCs) by the recruitment of the viral protein VP24. NCs are tightly coiled left-handed helices whose structure is mainly determined by the multimerization of NP at its N-terminus, and these helices form the inner layer of the NCs. The RNA genome is fixed by 2 lobes of the NP N-terminus and is thus guided by individual NP molecules along the turns of the helix. Direct interaction of the NP C-terminus with the VP35 and VP24 molecules forms the outer layer of the NCs. Once formed, NCs that are located at the border of the IBs recruit actin polymerization machinery to one of their ends to drive their transport to budding sites for their envelopment and final release. Here, we review the current knowledge on the structure, assembly, and transport of filovirus NCs.

Postural deficits such as hyperlordosis (hollow back) or hyperkyphosis (hunchback) are relevant health issues. Diagnoses depend on the experience of the examiner and are, therefore, often subjective and prone to errors. Machine learning (ML) methods in combination with explainable artificial intelligence (XAI) tools have proven useful for providing an objective, data-based orientation. However, only a few works have considered posture parameters, leaving the potential for more human-friendly XAI interpretations still untouched. Therefore, the present work proposes an objective, data-driven ML system for medical decision support that enables especially human-friendly interpretations using counterfactual explanations (CFs). The posture data for 1151 subjects were recorded by means of stereophotogrammetry. An expert-based classification of the subjects regarding the presence of hyperlordosis or hyperkyphosis was initially performed. Using a Gaussian progress classifier, the models were trained and interpreted using CFs. The label errors were flagged and re-evaluated using confident learning. Very good classification performances for both hyperlordosis and hyperkyphosis were found, whereby the re-evaluation and correction of the test labels led to a significant improvement (MPRAUC = 0.97). A statistical evaluation showed that the CFs seemed to be plausible, in general. In the context of personalized medicine, the present study’s approach could be of importance for reducing diagnostic errors and thereby improving the individual adaptation of therapeutic measures. Likewise, it could be a basis for the development of apps for preventive posture assessment.

Severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) utilizes cellular trafficking pathways to process its structural proteins and move them to the site of assembly. Nevertheless, the exact process of assembly and subcellular trafficking of SARS-CoV-2 proteins remains largely unknown. Here, we have identified and characterized Rab1B as an important host factor for the trafficking and maturation of the spike protein (S) after synthesis at the endoplasmic reticulum (ER). Using confocal microscopy, we showed that S and Rab1B substantially colocalized in compartments of the early secretory pathway. Co-expression of dominant-negative (DN) Rab1B N121I leads to an aberrant distribution of S into perinuclear spots after ectopic expression and in SARS-CoV-2-infected cells caused by either structural rearrangement of the ERGIC or Golgi or missing interaction between Rab1B and S. Western blot analyses revealed a complete loss of the mature, cleaved S2 subunit in cell lysates and culture supernatants upon co-expression of DN Rab1B N121I. In sum, our studies indicate that Rab1B is an important regulator of trafficking and maturation of SARS-CoV-2 S, which not only improves our understanding of the coronavirus replication cycle but also may have implications for the development of antiviral strategies.

The objectification of acute fatigue (during isometric muscle contraction) and cumulative fatigue (due to multiple intermittent isometric muscle contractions) plays an important role in sport climbing. The data of 42 participants were used in the study. Climbing performance was operationalized using maximal climbing-specific holding time (CSHT) by performing dead hangs. The test started with an initial measurement of handgrip strength (HGS) followed by three intermittent measurements of CSHT and HGS. During the test, finger flexor muscle oxygen saturation (SmO2) was measured using a near-infrared spectroscopy wearable biosensor. Significant reductions in CSHT and HGS could be found (p < 0.001), which indicates that the consecutive maximal isometric holding introduces cumulative fatigue. The reduction in CSHT did not correlate with a reduction in HGS over multiple consecutive maximal dead hangs (p > 0.35). Furthermore, there were no significant differences in initial SmO2 level, SmO2 level at termination, SmO2 recovery, and mean negative slope of the SmO2 saturation reduction between the different measurements (p > 0.24). Significant differences were found between pre-, termination-, and recovery- (10 s after termination) SmO2 levels (p < 0.001). Therefore, monitoring acute fatigue using athletes’ termination SmO2 saturation seems promising. By contrast, the measurement of HGS and muscle oxygen metabolism seems inappropriate for monitoring cumulative fatigue during intermittent isometric climbing-specific muscle contractions.

Summary: Preexisting immunity against SARS-CoV-2 may have critical implications for our understanding of COVID-19 susceptibility and severity. The presence and clinical relevance of a preexisting B cell immunity remain to be fully elucidated. Here, we provide a detailed analysis of the B cell immunity to SARS-CoV-2 in unexposed individuals. To this end, we extensively investigated SARS-CoV-2 humoral immunity in 150 adults sampled pre-pandemically. Comprehensive screening of donor plasma and purified IgG samples for binding and neutralization in various functional assays revealed no substantial activity against SARS-CoV-2 but broad reactivity to endemic betacoronaviruses. Moreover, we analyzed antibody sequences of 8,174 putatively SARS-CoV-2-reactive B cells at a single cell level and generated and tested 158 monoclonal antibodies. None of these antibodies displayed relevant binding or neutralizing activity against SARS-CoV-2. Taken together, our results show no evidence of competent preexisting antibody and B cell immunity against SARS-CoV-2 in unexposed adults.

Abstract Background Live-cell imaging is a powerful tool for visualization of the spatio-temporal dynamics of moving signals in living cells. Although this technique can be utilized to visualize nucleocapsid transport in Marburg virus (MARV)- or Ebola virus-infected cells, the experiments require biosafety level-4 (BSL-4) laboratories, which are restricted to trained and authorized individuals. Methods To overcome this limitation, we developed a live-cell imaging system to visualize MARV nucleocapsid-like structures using fluorescence-conjugated viral proteins, which can be conducted outside BSL-4 laboratories. Results Our experiments revealed that nucleocapsid-like structures have similar transport characteristics to those of nucleocapsids observed in MARV-infected cells, both of which are mediated by actin polymerization. Conclusions We developed a non-infectious live cell imaging system to visualize intracellular transport of MARV nucleocapsid-like structures. This system provides a safe platform to evaluate antiviral drugs that inhibit MARV nucleocapsid transport.

Ebola (EBOV) and Marburg virus (MARV) are highly pathogenic filoviruses that influence cellular signaling according to their own needs. MARV has been shown to regulate the IRE1α-dependent unfolded protein response (UPR) to ensure optimal virus replication. It was not known whether EBOV affects this signaling cascade, which can be beneficial or detrimental for viruses. Activation of IRE1α leads to the expression of the transcription factor XBP1s, which binds to cis-acting UPR elements (UPRE), resulting in the expression of genes aimed at restoring homeostasis in the endoplasmic reticulum. We observed that EBOV infection, in contrast to MARV infection, led to UPR activation by IRE1α-dependent but not ATF6-dependent signaling. We showed an activation of IRE1α, XBP1s and UPRE target genes upon EBOV infection. ATF6, another UPRE transcription factor, was not activated. UPRE activation was mainly attributed to the EBOV nucleoprotein NP and the soluble glycoprotein sGP. Finally, activation of UPR by thapsigargin, a potent ER-stress inducer, in parallel to infection as well as knock-out of XBP1 had no effect on EBOV growth, while MARV proliferation was affected by thapsigargin-dependent UPR activation. Taken together EBOV and MARV differ in their strategy of balancing IRE1α-dependent signaling for their own needs.

Transcription of non-segmented negative sense (NNS) RNA viruses follows a stop-start mechanism and is thought to be initiated at the genome’s very 3’-end. The synthesis of short abortive leader transcripts (leaderRNAs) has been linked to transcription initiation for some NNS viruses. Here, we identified the synthesis of abortive leaderRNAs (as well as trailer RNAs) that are specifically initiated opposite to (anti)genome nt 2; leaderRNAs are predominantly terminated in the region of nt ~ 60–80. LeaderRNA synthesis requires hexamer phasing in the 3’-leader promoter. We determined a steady-state NP mRNA:leaderRNA ratio of ~10 to 30-fold at 48 h after Ebola virus (EBOV) infection, and this ratio was higher (70 to 190-fold) for minigenome-transfected cells. LeaderRNA initiation at nt 2 and the range of termination sites were not affected by structure and length variation between promoter elements 1 and 2, nor the presence or absence of VP30. Synthesis of leaderRNA is suppressed in the presence of VP30 and termination of leaderRNA is not mediated by cryptic gene end (GE) signals in the 3’-leader promoter. We further found different genomic 3’-end nucleotide requirements for transcription versus replication, suggesting that promoter recognition is different in the replication and transcription mode of the EBOV polymerase. We further provide evidence arguing against a potential role of EBOV leaderRNAs as effector molecules in innate immunity. Taken together, our findings are consistent with a model according to which leaderRNAs are abortive replicative RNAs whose synthesis is not linked to transcription initiation. Rather, replication and transcription complexes are proposed to independently initiate RNA synthesis at separate sites in the 3’-leader promoter, i.e., at the second nucleotide of the genome 3’-end and at the more internally positioned transcription start site preceding the first gene, respectively, as reported for Vesicular stomatitis virus. Author summary The RNA polymerase (RdRp) of Ebola virus (EBOV) initiates RNA synthesis at the 3’-leader promoter of its encapsidated, non-segmented negative sense (NNS) RNA genome, either at the penultimate 3’-end position of the genome in the replicative mode or more internally (position 56) at the transcription start site (TSS) in its transcription mode. Here we identified the synthesis of abortive replicative RNAs that are specifically initiated opposite to genome nt 2 (termed leaderRNAs) and predominantly terminated in the region of nt ~ 60–80 near the TSS. The functional role of abortive leaderRNA synthesis is still enigmatic; a role in interferon induction could be excluded. Our findings indirectly link leaderRNA termination to nucleoprotein (NP) availability for encapsidation of nascent replicative RNA or to NP removal from the template RNA. Our findings further argue against the model that leaderRNA synthesis is a prerequisite for each transcription initiation event at the TSS. Rather, our findings are in line with the existence of distinct replicase and transcriptase complexes of RdRp that interact differently with the 3’-leader promoter and intiate RNA synthesis independently at different sites (position 2 or 56 of the genome), mechanistically similar to another NNS virus, Vesicular stomatitis virus.

Handgrip strength (HGS) appears to be an indicator of climbing performance. The transferability of HGS measurements obtained using a hand dynamometer and factors that influence the maximal climbing-specific holding time (CSHT) are largely unclear. Forty-eight healthy subjects (27 female, 21 male; age: 22.46 ± 3.17 years; height: 172.76 ± 8.91 cm; weight: 69.07 ± 12.41 kg; body fat: 20.05% ± 7.95%) underwent a maximal pull-up test prior to the experiment and completed a self-assessment using a Likert scale questionnaire. HGS was measured using a hand dynamometer, whereas CSHT was measured using a fingerboard. Multiple linear regressions showed that weight, maximal number of pull-ups, HGS normalized by subject weight, and length of the middle finger had a significant effect on the maximal CSHT (non-dominant hand: R2corr = 0.63; dominant hand: R2corr = 0.55). Deeper exploration using a machine learning model including all available data showed a predictive performance with R2 = 0.51 and identified another relevant parameter for the regression model. These results call into question the use of hand dynamometers and highlight the performance-related importance of body weight in climbing practice. The results provide initial indications that finger length may be used as a sub-factor in talent scouting.

Ebola virus is the causative agent of Ebola virus disease, a severe, often fatal illness in humans. So far, there are no US Food and Drug Administration (FDA)-approved therapeutics directed against Ebola virus. Here, we selected the host factor Niemann-Pick C1 (NPC1), which has been shown to be essential for Ebola virus entry into host cytoplasm, as a therapeutic target for suppression by locked nucleic acid-modified antisense oligonucleotides. Screening of antisense oligonucleotides in human and murine cell lines led to identification of candidates with up to 94% knockdown efficiency and 50% inhibitory concentration (IC50) values in the submicromolar range. Selected candidate oligonucleotides led to efficient NPC1 protein knockdown in vitro without alteration of cell viability. Furthermore, they did not have immune stimulatory activity in cell-based assays. Treatment of Ebola-virus-infected HeLa cells with the most promising candidates resulted in significant (>99%) virus titer reduction, indicating that antisense oligonucleotides against NPC1 are a promising therapeutic approach for treatment of Ebola virus infection. Keywords: Ebola virus, antisense oligonucleotide, NPC1, LNA

ABSTRACTViruses regulate cellular signalling pathways to ensure optimal viral replication. During Marburg virus (MARV) infection, large quantities of the viral glycoprotein GP are produced in the ER; this may result in the activation of the unfolded protein response (UPR). The most conserved pathway to trigger UPR is initiated by IRE1. Activation of IRE1 results in auto-phosphorylation, splicing of the XBP1 mRNA and translation of the XBP1s protein. XBP1s binds cis-acting UPR elements (UPRE) which leads to the enhanced expression of genes which should restore ER homeostasis. XBP1u protein is translated, if IRE1 is not activated. Here we show that ectopic expression of MARV GP activated the IRE1-XBP1 axis of UPR as monitored by UPRE luciferase assays. However, while at 24 h of infection with MARV IRE1 was phosphorylated, expression of XBP1s was only slightly enhanced and UPRE activity was not detected. The IRE1-XBP1 axis was not active at 48 h p.i. Co-expression studies of MARV proteins demonstrated that the MARV protein VP30 suppressed UPRE activation. Co-immunoprecipitation analyses revealed an RNA-dependent interaction of VP30 with XBP1u. Knock-out of IRE1 supported MARV infection at late time points. Taken together, these results suggest that efficient MARV propagation requires specific regulation of IRE1 activity.

Summary: Ebola virus (EBOV) is responsible for outbreaks with case fatality rates of up to 90% and for an epidemic in West Africa with more than ten thousand deaths. EBOV glycoprotein (EBOV-GP) is the only viral surface protein and is responsible for viral entry into cells. Here, by employing pseudotyped EBOV-GP viral particles, we uncover a critical role for sphingolipids in inhibiting viral entry. Sphingosine kinase 1 (SphK1) catalyzes the phosphorylation of sphingosine to sphingosine 1-phosphate (S1P). The administration of the SphK1 activator, K6PC-5, or S1P, or the overexpression of SphK1 consistently exhibited striking inhibitory effects in EBOV-GP-driven entry in diverse cell lines. Finally, K6PC-5 markedly reduced the EBOV titer in infected cells and the de novo production of viral proteins. These data present K6PC-5 as an efficient tool to inhibit EBOV infection in endothelial cells and suggest further studies to evaluate its systemic effects.

The urgent need for vaccines against Ebola virus (EBOV) was underscored by the large outbreak in West Africa (2014–2016). Since then, several promising vaccine candidates have been tested in pre-clinical and clinical studies. As a result, two vaccines were approved for human use in 2019/2020, of which one includes a heterologous adenovirus/Modified Vaccinia virus Ankara (MVA) prime-boost regimen. Here, we tested new vaccine candidates based on the recombinant MVA vector, encoding the EBOV nucleoprotein (MVA-EBOV-NP) or glycoprotein (MVA-EBOV-GP) for their efficacy after homologous prime-boost immunization in mice. Our aim was to investigate the role of each antigen in terms of efficacy and correlates of protection. Sera of mice vaccinated with MVA-EBOV-GP were virus-neutralizing and MVA-EBOV-NP immunization readily elicited interferon-γ-producing NP-specific CD8+ T cells. While mock-vaccinated mice succumbed to EBOV infection, all vaccinated mice survived and showed drastically decreased viral loads in sera and organs. In addition, MVA-EBOV-NP vaccinated mice became susceptible to lethal EBOV infection after depletion of CD8+ T cells prior to challenge. This study highlights the potential of MVA-based vaccines to elicit humoral immune responses as well as a strong and protective CD8+ T cell response and contributes to understanding the possible underlying mechanisms.

Filoviruses such as Ebola and Marburg virus bud from the host membrane as enveloped virions. This process is achieved by the matrix protein VP40. When expressed alone, VP40 induces budding of filamentous virus-like particles, suggesting that localization to the plasma membrane, oligomerization into a matrix layer, and generation of membrane curvature are intrinsic properties of VP40. There has been no direct information on the structure of VP40 matrix layers within viruses or virus-like particles. We present structures of Ebola and Marburg VP40 matrix layers in intact virus-like particles, and within intact Marburg viruses. VP40 dimers assemble extended chains via C-terminal domain interactions. These chains stack to form 2D matrix lattices below the membrane surface. These lattices form a patchwork assembly across the membrane and suggesting that assembly may begin at multiple points. Our observations define the structure and arrangement of the matrix protein layer that mediates formation of filovirus particles.

ABSTRACT Ebola virus (EBOV) causes a severe and often fatal disease for which no approved vaccines or antivirals are currently available. EBOV VP30 has been described as a viral phosphoprotein, and nonphosphorylated VP30 is essential and sufficient to support secondary transcription in an EBOV-specific minigenome system; however, phosphorylatable serine residues near the N terminus of VP30 are required to support primary viral transcription as well as the reinitiation of VP30-mediated transcription at internal EBOV genes. While the dephosphorylation of VP30 by the cellular phosphatase PP2A was found to be mediated by nucleoprotein, the VP30-specific kinases and the role of phosphorylation remain unknown. Here, we report that serine-arginine protein kinase 1 (SRPK1) and SRPK2 phosphorylate serine 29 of VP30, which is located in an N-terminal R26xxS29 motif. Interaction with VP30 via the R26xxS29 motif recruits SRPK1 into EBOV-induced inclusion bodies, the sites of viral RNA synthesis, and an inhibitor of SRPK1/SRPK2 downregulates primary viral transcription. When the SRPK1 recognition motif of VP30 was mutated in a recombinant EBOV, virus replication was severely impaired. It is presumed that the interplay between SRPK1 and PP2A in the EBOV inclusions provides a comprehensive regulatory circuit to ensure the activity of VP30 in EBOV transcription. Thus, the identification of SRPK1 is an important mosaic stone that completes our picture of the players involved in Ebola virus transcription regulation. IMPORTANCE The largest Ebola virus (EBOV) epidemic in West Africa ever caused more than 28,000 cases and 11,000 deaths, and the current EBOV epidemic in the Democratic Republic of the Congo continues, with more than 3,000 cases to date. Therefore, it is essential to develop antivirals against EBOV. Recently, an inhibitor of the cellular phosphatase PP2A-mediated dephosphorylation of the EBOV transcription factor VP30 has been shown to suppress the spread of Ebola virus. Here, we identified the protein kinase SRPK1 as a VP30-specific kinase that phosphorylates serine 29, the same residue that is dephosphorylated by PP2A. SRPK1-mediated phosphorylation of serine 29 enabled primary viral transcription. Mutation of the SRPK1 recognition motif in VP30 resulted in significant growth inhibition of EBOV. Similarly, elevation of the phosphorylation status of serine 29 by overexpression of SRPK1 inhibited EBOV growth, highlighting the importance of reversible phosphorylation of VP30 as a potential therapeutic target.

We describe a strain of Lassa virus representing a putative new lineage that was isolated from a cluster of human infections with an epidemiologic link to Togo. This finding extends the known range of Lassa virus to Togo.

Despite the recent availability of vaccines against severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2), there is an urgent need for specific anti-SARS-CoV-2 drugs. Monoclonal neutralizing antibodies are an important drug class in the global fight against the SARS-CoV-2 pandemic due to their ability to convey immediate protection and their potential to be used as both prophylactic and therapeutic drugs. Clinically used neutralizing antibodies against respiratory viruses are currently injected intravenously, which can lead to suboptimal pulmonary bioavailability and thus to a lower effectiveness. Here we describe DZIF-10c, a fully human monoclonal neutralizing antibody that binds the receptor-binding domain of the SARS-CoV-2 spike protein. DZIF-10c displays an exceptionally high neutralizing potency against SARS-CoV-2, retains full activity against the variant of concern (VOC) B.1.1.7 and still neutralizes the VOC B.1.351, although with reduced potency. Importantly, not only systemic but also intranasal application of DZIF-10c abolished the presence of infectious particles in the lungs of SARS-CoV-2 infected mice and mitigated lung pathology when administered prophylactically. Along with a favorable pharmacokinetic profile, these results highlight DZIF-10c as a novel human SARS-CoV-2 neutralizing antibody with high in vitro and in vivo antiviral potency. The successful intranasal application of DZIF-10c paves the way for clinical trials investigating topical delivery of anti-SARS-CoV-2 antibodies.

Predicting vaccine efficacy remains a challenge. We used a systems vaccinology approach to identify early innate immune correlates of antibody induction in humans receiving the Ebola vaccine rVSV-ZEBOV. Blood samples from days 0, 1, 3, 7, and 14 were analyzed for changes in cytokine levels, innate immune cell subsets, and gene expression. Integrative statistical analyses with cross-validation identified a signature of 5 early innate markers correlating with antibody titers on day 28 and beyond. Among those, IP-10 on day 3 and MFI of CXCR6 on NK cells on day 1 were independent correlates. Consistently, we found an early gene expression signature linked to IP-10. This comprehensive characterization of early innate immune responses to the rVSV-ZEBOV vaccine in humans revealed immune signatures linked to IP-10. These results suggest correlates of vaccine-induced antibody induction and provide a rationale to explore strategies for augmenting the effectiveness of vaccines through manipulation of IP-10.

Abstract Background The ideal protein expression system should provide recombinant proteins in high quality and quantity involving low production costs only. However, especially for complex therapeutic proteins like monoclonal antibodies many challenges remain to meet this goal and up to now production of monoclonal antibodies is very costly and delicate. Particularly, emerging disease outbreaks like Ebola virus in Western Africa in 2014–2016 make it necessary to reevaluate existing production platforms and develop robust and cheap alternatives that are easy to handle. Results In this study, we engineered the microalga Phaeodactylum tricornutum to produce monoclonal IgG antibodies against the nucleoprotein of Marburg virus, a close relative of Ebola virus causing severe hemorrhagic fever with high fatality rates in humans. Sequences for both chains of a mouse IgG antibody were retrieved from a murine hybridoma cell line and implemented in the microalgal system. Fully assembled antibodies were shown to be secreted by the alga and antibodies were proven to be functional in western blot, ELISA as well as IFA studies just like the original hybridoma produced IgG. Furthermore, synthetic variants with constant regions of a rabbit IgG and human IgG with optimized codon usage were produced and characterized. Conclusions This study highlights the potential of microalgae as robust and low cost expression platform for monoclonal antibodies secreting IgG antibodies directly into the culture medium. Microalgae possess rapid growth rates, need basically only water, air and sunlight for cultivation and are very easy to handle.

Background: The recent West African Ebola epidemic led to accelerated efforts to test Ebola vaccine candidates. As part of the World Health Organisation-led VSV Ebola Consortium (VEBCON), we performed a phase I clinical trial investigating rVSV-ZEBOV (a recombinant vesicular stomatitis virus-vectored Ebola vaccine), which has recently demonstrated protection from Ebola virus disease (EVD) in phase III clinical trials and is currently in advanced stages of licensing. So far, correlates of immune protection are incompletely understood and the role of cell-mediated immune responses has not been comprehensively investigated to date. Methods: We recruited 30 healthy subjects aged 18–55 into an open-label, dose-escalation phase I trial testing three doses of rVSV-ZEBOV (3 × 105 plaque-forming units (PFU), 3 × 106 PFU, 2 × 107 PFU) (ClinicalTrials.gov; NCT02283099). Main study objectives were safety and immunogenicity, while exploratory objectives included lymphocyte dynamics, cell-mediated immunity and cytokine networks, which were assessed using flow cytometry, ELISpot and LUMINEX assay. Findings: Immunization with rVSV-ZEBOV was well tolerated without serious vaccine-related adverse events. Ebola virus-specific neutralizing antibodies were induced in nearly all individuals. Additionally, vaccinees, particularly within the highest dose cohort, generated Ebola glycoprotein (GP)-specific T cells and initiated a cascade of signaling molecules following stimulation of peripheral blood mononuclear cells with Ebola GP peptides. Interpretation: In addition to a benign safety and robust humoral immunogenicity profile, subjects immunized with 2 × 107 PFU elicited higher cellular immune responses and stronger interlocked cytokine networks compared to lower dose groups. To our knowledge these data represent the first detailed cell-mediated immuneprofile of a clinical trial testing rVSV-ZEBOV, which is of particular interest in light of its potential upcoming licensure as the first Ebola vaccine. VEBCON trial Hamburg, Germany (NCT02283099).

Muscular imbalances of the trunk muscles are held responsible for changes in body posture. At the same time, whole-body electromyostimulation (WB-EMS) has been established as a new training method that enables simultaneous stimulation of many muscle groups. This study was aiming to analyze if a 10 weeks WB-EMS training changes posture-relevant parameters and/or improves isometric strength of the trunk extensors and flexors, and if there are differences based on stimulation at 20 Hz and 85 Hz. Fifty eight untrained adult test persons were divided into three groups (control, CON; training with 20 Hz stimulation, TR20; training with 85 Hz, TR85). Anthropometric parameters, trunk extension and flexion forces and torques, and posture parameters were determined before (n = 58) and after (n = 53: CON: n = 15, TR20: n = 19, TR85: n = 19) a 10 weeks WB-EMS training program (15 applications, 9 exercises). Differences between the groups were calculated for pre- and post-tests using univariate ANOVA and between the test times using repeated (2 × 3) ANOVA. Comparisons of pairs were calculated post hoc based on Fisher (LSD). No differences between the groups were found for the posture parameters. The post hoc analysis of both trunk flexion and trunk extension forces and torques showed a significant difference between the groups TR85 and CON but no difference between the other group pairs. A 10 weeks whole-body electrostimulation training with a stimulation frequency of 85 Hz in contrast to training with a stimulation frequency of 20 Hz improves the trunk muscle strength of an untrained group but does not significantly change posture parameters.

Intensity regulation during whole-body electromyostimulation (WB-EMS) training is mostly controlled by subjective scales such as CR-10 Borg scale. To determine objective training intensities derived from a maximum as it is used in conventional strength training using the one-repetition-maximum (1-RM), a comparable maximum in WB-EMS is necessary. Therefore, the aim of this study was to examine, if there is an individual maximum intensity tolerance plateau after multiple consecutive EMS application sessions. A total of 52 subjects (24.1 ± 3.2 years; 76.8 ± 11.1 kg; 1.77 ± 0.09 m) participated in the longitudinal, observational study (38 males, 14 females). Each participant carried out four consecutive maximal EMS applications (T1–T4) separated by 1 week. All muscle groups were stimulated successively until their individual maximum and combined to a whole-body stimulation index to carry out a possible statement for the development of the maximum intensity tolerance of the whole body. There was a significant main effect between the measurement times for all participants (p < 0.001; 𝜂2 = 0.39) as well as gender specific for males (p = 0.001; 𝜂2 = 0.18) and females (p < 0.001; 𝜂2 = 0.57). There were no interaction effects of gender × measurement time (p = 0.394). The maximum intensity tolerance increased significantly from T1 to T2 (p = 0.001) and T2 to T3 (p < 0.001). There was no significant difference between T3 and T4 (p = 1.0). These results indicate that there is an adjustment of the individual maximum intensity tolerance to a WB-EMS training after three consecutive tests. Therefore, there is a need of several habituation units comparable to the identification of the individual 1-RM in conventional strength training. Further research should focus on an objective intensity-specific regulation of the WB-EMS based on the individual maximum intensity tolerance to characterize different training areas and therefore generate specific adaptations to a WB-EMS training compared to conventional strength training methods.

A common problem in dolomite reservoirs is the heterogeneous distribution of porosity-reducing diagenetic phases. The intrasalt carbonates of the Ediacaran-Early Cambrian Ara Group in the South Oman Salt Basin represent a self-sourcing petroleum system. Depositional facies and carbonate/evaporite platform architecture are well understood, but original reservoir properties have been modified by diagenesis. Some of the carbonate reservoirs failed to produce hydrocarbons at acceptable rates, which triggered this study. The extent of primary porosity reduction by diagenetic phases was quantified using point counting. To visualize the distribution of diagenetic phases on a field scale, we constructed 2D interpolation diagenesis maps to identify patterns in cementation. The relative timing of diagenetic events was constrained based on thin-section observations and stable isotope analyses. Near-surface diagenesis is dominated by reflux-related processes, leading to porosity inversion in initial highly porous facies and a patchy distribution of early cements. This strong diagenetic overprint of primary and early diagenetic porosity by reflux-related cements leads to a reduction of stratigraphic and facies control on porosity. Calcite was identified as a burial-related cement phase that leads to an almost complete loss of intercrystalline porosity and permeability. Bitumen is an important pore-occluding phase and time marker of the deep-burial realm. The stratigraphic position of the dolomite reservoirs embedded at the base of a salt diapir had a strong impact on its diagenetic development. The salt isolated the dolomites from external fluids, leading to a closed system diagenesis and the buildup of near lithostatic fluid pressures. In combination, these processes decreased the impact of further burial diagenetic processes. The study highlights that cement distribution in salt-encased carbonate reservoirs is mainly related to early diagenetic processes but can be very heterogeneous on a field scale. Further work is needed to implement these heterogeneities in an integrated numerical reservoir model.

Background: The positive effect of carbohydrates from commercial beverages on soccer-specific exercise has been clearly demonstrated. However, no study is available that uses a home-mixed beverage in a test where technical skills were required. Methods: Nine subjects participated voluntarily in this double-blind, randomized, placebo-controlled crossover study. On three testing days, the subjects performed six Hoff tests with a 3-min active break as a preload and then the Yo-Yo Intermittent Running Test Level 1 (Yo-Yo IR1) until exhaustion. On test days 2 and 3, the subjects received either a 69 g carbohydrate-containing drink (syrup–water mixture) or a carbohydrate-free drink (aromatic water). Beverages were given in several doses of 250 mL each: 30 min before and immediately before the exercise and after 18 and 39 min of exercise. The primary target parameters were the running performance in the Hoff test and Yo-Yo IR1, body mass and heart rate. Statistical differences between the variables of both conditions were analyzed using paired samples t-tests. Results: The maximum heart rate in Yo-Yo IR1 showed significant differences (syrup: 191.1 ± 6.2 bpm; placebo: 188.0 ± 6.89 bpm; t(6) = −2.556; p = 0.043; dz = 0.97). The running performance in Yo-Yo IR1 under the condition syrup significantly increased by 93.33 ± 84.85 m (0–240 m) on average (p = 0.011). Conclusions: The intake of a syrup–water mixture with a total of 69 g carbohydrates leads to an increase in high-intensive running performance after soccer specific loads. Therefore, the intake of carbohydrate solutions is recommended for intermittent loads and should be increasingly considered by coaches and players.

The intracellular transport of nucleocapsids of the highly pathogenic Marburg, as well as Ebola virus (MARV, EBOV), represents a critical step during the viral life cycle. Intriguingly, a population of these nucleocapsids is distributed over long distances in a directed and polar fashion. Recently, it has been demonstrated that the intracellular transport of filoviral nucleocapsids depends on actin polymerization. While it was shown that EBOV requires Arp2/3-dependent actin dynamics, the details of how the virus exploits host actin signaling during intracellular transport are largely unknown. Here, we apply a minimalistic transfection system to follow the nucleocapsid-like structures (NCLS) in living cells, which can be used to robustly quantify NCLS transport in live cell imaging experiments. Furthermore, in cells co-expressing LifeAct, a marker for actin dynamics, NCLS transport is accompanied by pulsative actin tails appearing on the rear end of NCLS. These actin tails can also be preserved in fixed cells, and can be visualized via high resolution imaging using STORM in transfected, as well as EBOV infected, cells. The application of inhibitory drugs and siRNA depletion against actin regulators indicated that EBOV NCLS utilize the canonical Arp2/3-Wave1-Rac1 pathway for long-distance transport in cells. These findings highlight the relevance of the regulation of actin polymerization during directed EBOV nucleocapsid transport in human cells.

BackgroundThe rVSVΔG-ZEBOV-GP vaccine prevented Ebola virus disease when used at 2 × 107 plaque-forming units (PFU) in a trial in Guinea. This study provides further safety and immunogenicity data.Methods and findingsA randomised, open-label phase I trial in Lambaréné, Gabon, studied 5 single intramuscular vaccine doses of 3 × 103, 3 × 104, 3 × 105, 3 × 106, or 2 × 107 PFU in 115 adults and a dose of 2 × 107 PFU in 20 adolescents and 20 children. The primary objective was safety and tolerability 28 days post-injection. Immunogenicity, viraemia, and shedding post-vaccination were evaluated as secondary objectives. In adults, mild-to-moderate adverse events were frequent, but there were no serious or severe adverse events related to vaccination. Before vaccination, Zaire Ebola virus (ZEBOV)-glycoprotein (GP)-specific and ZEBOV antibodies were detected in 11% and 27% of adults, respectively. In adults, 74%-100% of individuals who received a dose 3 × 104, 3 × 105, 3 × 106, or 2 × 107 PFU had a ≥4.0-fold increase in geometric mean titres (GMTs) of ZEBOV-GP-specific antibodies at day 28, reaching GMTs of 489 (95% CI: 264-908), 556 (95% CI: 280-1,101), 1,245 (95% CI: 899-1,724), and 1,503 (95% CI: 931-2,426), respectively. Twenty-two percent of adults had a ≥4-fold increase of ZEBOV antibodies, with GMTs at day 28 of 1,015 (647-1,591), 1,887 (1,154-3,085), 1,445 (1,013-2,062), and 3,958 (2,249-6,967) for the same doses, respectively. These antibodies persisted up to day 180 for doses ≥3 × 105 PFU. Adults with antibodies before vaccination had higher GMTs throughout. Neutralising antibodies were detected in more than 50% of participants at doses ≥3 × 105 PFU. As in adults, no serious or severe adverse events related to vaccine occurred in adolescents or children. At day 2, vaccine RNA titres were higher for adolescents and children than adults. At day 7, 78% of adolescents and 35% of children had recombinant vesicular stomatitis virus RNA detectable in saliva. The vaccine induced high GMTs of ZEBOV-GP-specific antibodies at day 28 in adolescents, 1,428 (95% CI: 1,025-1,989), and children, 1,620 (95% CI: 806-3,259), and in both groups antibody titres increased up to day 180. The absence of a control group, lack of stratification for baseline antibody status, and imbalances in male/female ratio are the main limitations of this study.ConclusionsOur data confirm the acceptable safety and immunogenicity profile of the 2 × 107 PFU dose in adults and support consideration of lower doses for paediatric populations and those who request boosting.Trial registrationPan African Clinical Trials Registry PACTR201411000919191.

During the recent Ebola outbreak in West Africa several international mobile laboratories were deployed to the mainly affected countries Guinea, Sierra Leone and Liberia to provide ebolavirus diagnostic capacity. Additionally, imported cases and small outbreaks in other countries required global preparedness for Ebola diagnostics. Detection of viral RNA by reverse transcription polymerase chain reaction has proven effective for diagnosis of ebolavirus disease and several assays are available. However, reliability of these assays is largely unknown and requires serious evaluation. Therefore, a proficiency test panel of 11 samples was generated and distributed on a global scale. Panels were analyzed by 83 expert laboratories and 106 data sets were returned. From these 78 results were rated optimal and 3 acceptable, 25 indicated need for improvement. While performance of the laboratories deployed to West Africa was superior to the overall performance there was no significant difference between the different assays applied.

Pneumonia may be caused by a wide range of pathogens and is considered the most common infectious cause of death in humans. Murine acute lung infection models mirror human pathologies in many aspects and contribute to our understanding of the disease and the development of novel treatment strategies. Despite progress in other fields of tissue imaging, histopathology remains the most conclusive and practical read out tool for the descriptive and semiquantitative evaluation of mouse pneumonia and therapeutic interventions. Here, we systematically describe and compare the distinctive histopathological features of established models of acute pneumonia in mice induced by Streptococcus (S.) pneumoniae, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Legionella pneumophila, Escherichia coli, Middle East respiratory syndrome (MERS) coronavirus, influenza A virus (IAV) and superinfection of IAV-incuced pneumonia with S. pneumoniae. Systematic comparisons of the models revealed striking differences in the distribution of lesions, the characteristics of pneumonia induced, principal inflammatory cell types, lesions in adjacent tissues, and the detectability of the pathogens in histological sections. We therefore identified core criteria for each model suitable for practical semiquantitative scoring systems that take into account the pathogen- and model-specific patterns of pneumonia. Other critical factors that affect experimental pathologies are discussed, including infectious dose, time kinetics, and the genetic background of the mouse strain. The substantial differences between the model-specific pathologies underscore the necessity of pathogen- and model-adapted criteria for the comparative quantification of experimental outcomes. These criteria also allow for the standardized validation and comparison of treatment strategies in preclinical models.

Sequence determination of complete or coding-complete genomes of viruses is becoming common practice for supporting the work of epidemiologists, ecologists, virologists, and taxonomists. Sequencing duration and costs are rapidly decreasing, sequencing hardware is under modification for use by non-experts, and software is constantly being improved to simplify sequence data management and analysis. Thus, analysis of virus disease outbreaks on the molecular level is now feasible, including characterization of the evolution of individual virus populations in single patients over time. The increasing accumulation of sequencing data creates a management problem for the curators of commonly used sequence databases and an entry retrieval problem for end users. Therefore, utilizing the data to their fullest potential will require setting nomenclature and annotation standards for virus isolates and associated genomic sequences. The National Center for Biotechnology Information’s (NCBI’s) RefSeq is a non-redundant, curated database for reference (or type) nucleotide sequence records that supplies source data to numerous other databases. Building on recently proposed templates for filovirus variant naming [ ()////-], we report consensus decisions from a majority of past and currently active filovirus experts on the eight filovirus type variants and isolates to be represented in RefSeq, their final designations, and their associated sequences.

The host cell protein tetherin can restrict the release of enveloped viruses from infected cells. The HIV-1 protein Vpu counteracts tetherin by removing it from the site of viral budding, the plasma membrane, and this process depends on specific interactions between the transmembrane domains of Vpu and tetherin. In contrast, the glycoproteins (GPs) of two filoviruses, Ebola and Marburg virus, antagonize tetherin without reducing surface expression, and the domains in GP required for tetherin counteraction are unknown. Here, we show that filovirus GPs depend on the presence of their authentic transmembrane domains for virus-cell fusion and tetherin antagonism. However, conserved residues within the transmembrane domain were dispensable for membrane fusion and tetherin counteraction. Moreover, the insertion of the transmembrane domain into a heterologous viral GP, Lassa virus GPC, was not sufficient to confer tetherin antagonism to the recipient. Finally, mutation of conserved residues within the fusion peptide of Ebola virus GP inhibited virus-cell fusion but did not ablate tetherin counteraction, indicating that the fusion peptide and the ability of GP to drive host cell entry are not required for tetherin counteraction. These results suggest that the transmembrane domains of filoviral GPs contribute to tetherin antagonism but are not the sole determinants.

(1) Background: The importance of a strong and stable trunk musculature is well known, but there is a lack of reliable, valid and objective test batteries with the necessary test economy, practicability and cost-benefit ratio. The aim of the present study was to evaluate the Bourban test for the validity of its exercise selection representing the ventral, right/left lateral and dorsal muscle chain. (2) Methods: 33 male sports students (24.1 ± 2.4 years, 181.6 ± 5.5 cm, 80.8 ± 7.3 kg) participated in the study. Median Frequency (MDF) and Mean Frequency (MNF) were calculated from the electromyographic signals and used to check whether fatigue of the suggested target musculature actually occurs during the different exercises and thus the exercise is representative for this part of the trunk. (3) Results: In all exercises significant fatigue was measured for MDF and MNF in the muscles working as agonists. (4) Conclusion: It can be stated that the Bourban trunk muscle strength test is a valid and economic test instrument for the evaluation of trunk strength (endurance). Compared to technically supported measuring systems, the Bourban test seems to be a flexible and cost-effective alternative for the broad mass.