@article{JafarnezhadgeroNorooziFakhrietal.2022, author = {Jafarnezhadgero, Amir Ali and Noroozi, Raha and Fakhri, Ehsan and Granacher, Urs and Oliveira, Anderson Souza}, title = {The Impact of COVID-19 and muscle fatigue on cardiorespiratory fitness and running kinetics in female recreational runners}, series = {Frontiers in physiology}, volume = {13}, journal = {Frontiers in physiology}, publisher = {Frontiers Media}, address = {Lausanne}, issn = {1664-042X}, doi = {10.3389/fphys.2022.942589}, pages = {10}, year = {2022}, abstract = {Background: There is evidence that fully recovered COVID-19 patients usually resume physical exercise, but do not perform at the same intensity level performed prior to infection. The aim of this study was to evaluate the impact of COVID-19 infection and recovery as well as muscle fatigue on cardiorespiratory fitness and running biomechanics in female recreational runners. Methods: Twenty-eight females were divided into a group of hospitalized and recovered COVID-19 patients (COV, n = 14, at least 14 days following recovery) and a group of healthy age-matched controls (CTR, n = 14). Ground reaction forces from stepping on a force plate while barefoot overground running at 3.3 m/s was measured before and after a fatiguing protocol. The fatigue protocol consisted of incrementally increasing running speed until reaching a score of 13 on the 6-20 Borg scale, followed by steady-state running until exhaustion. The effects of group and fatigue were assessed for steady-state running duration, steady-state running speed, ground contact time, vertical instantaneous loading rate and peak propulsion force. Results: COV runners completed only 56\% of the running time achieved by the CTR (p < 0.0001), and at a 26\% slower steady-state running speed (p < 0.0001). There were fatigue-related reductions in loading rate (p = 0.004) without group differences. Increased ground contact time (p = 0.002) and reduced peak propulsion force (p = 0.005) were found for COV when compared to CTR. Conclusion: Our results suggest that female runners who recovered from COVID-19 showed compromised running endurance and altered running kinetics in the form of longer stance periods and weaker propulsion forces. More research is needed in this area using larger sample sizes to confirm our study findings.}, language = {en} } @article{PetazziKoikkarahAjiTischleretal.2021, author = {Petazzi, Roberto Arturo and Koikkarah Aji, Amit and Tischler, Nicole D. and Chiantia, Salvatore}, title = {Detection of envelope glycoprotein assembly from old world hantaviruses in the Golgi apparatus of living cells}, series = {Journal of virology}, volume = {95}, journal = {Journal of virology}, number = {4}, publisher = {American Society for Microbiology}, address = {Baltimore, Md.}, issn = {1098-5514}, doi = {10.1128/JVI.01238-20}, pages = {18}, year = {2021}, abstract = {Hantaviruses are emerging pathogens that occasionally cause deadly outbreaks in the human population. While the structure of the viral envelope has been characterized with high precision, protein-protein interactions leading to the formation of new virions in infected cells are not fully understood. We used quantitative fluorescence microscopy (i.e., number and brightness analysis and fluorescence fluctuation spectroscopy) to monitor the interactions that lead to oligomeric spike complex formation in the physiological context of living cells. To this aim, we quantified protein-protein interactions for the glycoproteins Gn and Gc from Puumala and Hantaan orthohantaviruses in several cellular models. The oligomerization of each protein was analyzed in relation to subcellular localization, concentration, and the concentration of its interaction partner. Our results indicate that, when expressed separately, Gn and Gc form, respectively, homo-tetrameric and homo-dimeric complexes, in a concentration-dependent manner. Site-directed mutations or deletion mutants showed the specificity of their homotypic interactions. When both glycoproteins were coexpressed, we observed in the Golgi apparatus clear indication of GnGc interactions and the formation of Gn-Gc multimeric protein complexes of different sizes, while using various labeling schemes to minimize the influence of the fluorescent tags. Such large glycoprotein multimers may be identified as multiple Gn viral spikes interconnected via Gc-Gc contacts. This observation provides the possible first evidence for the initial assembly steps of the viral envelope within this organelle, and does so directly in living cells.
IMPORTANCE In this work, we investigate protein-protein interactions that drive the assembly of the hantavirus envelope. These emerging pathogens have the potential to cause deadly outbreaks in the human population. Therefore, it is important to improve our quantitative understanding of the viral assembly process in infected cells, from a molecular point of view. By applying advanced fluorescence microscopy methods, we monitored the formation of viral spike complexes in different cell types. Our data support a model for hantavirus assembly according to which viral spikes are formed via the clustering of hetero-dimers of the two viral glycoproteins Gn and Gc. Furthermore, the observation of large Gn-Gc hetero-multimers provide the possible first evidence for the initial assembly steps of the viral envelope, directly in the Golgi apparatus of living cells.}, language = {en} } @phdthesis{Stanke2023, author = {Stanke, Sandra}, title = {AC electrokinetic immobilization of influenza viruses and antibodies on nanoelectrode arrays for on-chip immunoassays}, doi = {10.25932/publishup-61716}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-617165}, school = {Universit{\"a}t Potsdam}, pages = {x, 115}, year = {2023}, abstract = {In the present thesis, AC electrokinetic forces, like dielectrophoresis and AC electroosmosis, were demonstrated as a simple and fast method to functionalize the surface of nanoelectrodes with submicrometer sized biological objects. These nanoelectrodes have a cylindrical shape with a diameter of 500 nm arranged in an array of 6256 electrodes. Due to its medical relevance influenza virus as well as anti-influenza antibodies were chosen as a model organism. Common methods to bring antibodies or proteins to biosensor surfaces are complex and time-consuming. In the present work, it was demonstrated that by applying AC electric fields influenza viruses and antibodies can be immobilized onto the nanoelectrodes within seconds without any prior chemical modification of neither the surface nor the immobilized biological object. The distribution of these immobilized objects is not uniform over the entire array, it exhibits a decreasing gradient from the outer row to the inner ones. Different causes for this gradient have been discussed, such as the vortex-shaped fluid motion above the nanoelectrodes generated by, among others, electrothermal fluid flow. It was demonstrated that parts of the accumulated material are permanently immobilized to the electrodes. This is a unique characteristic of the presented system since in the literature the AC electrokinetic immobilization is almost entirely presented as a method just for temporary immobilization. The spatial distribution of the immobilized viral material or the anti-influenza antibodies at the electrodes was observed by either the combination of fluorescence microscopy and deconvolution or by super-resolution microscopy (STED). On-chip immunoassays were performed to examine the suitability of the functionalized electrodes as a potential affinity-based biosensor. Two approaches were pursued: A) the influenza virus as the bio-receptor or B) the influenza virus as the analyte. Different sources of error were eliminated by ELISA and passivation experiments. Hence, the activity of the immobilized object was inspected by incubation with the analyte. This resulted in the successful detection of anti-influenza antibodies by the immobilized viral material. On the other hand, a detection of influenza virus particles by the immobilized anti-influenza antibodies was not possible. The latter might be due to lost activity or wrong orientation of the antibodies. Thus, further examinations on the activity of by AC electric fields immobilized antibodies should follow. When combined with microfluidics and an electrical read-out system, the functionalized chips possess the potential to serve as a rapid, portable, and cost-effective point-of-care (POC) device. This device can be utilized as a basis for diverse applications in diagnosing and treating influenza, as well as various other pathogens.}, language = {en} } @phdthesis{Kruse2023, author = {Kruse, Marlen}, title = {Characterization of biomolecules and their interactions using electrically controllable DNA nanolevers}, doi = {10.25932/publishup-57738}, url = {http://nbn-resolving.de/urn:nbn:de:kobv:517-opus4-577384}, school = {Universit{\"a}t Potsdam}, pages = {100, xxii}, year = {2023}, abstract = {In this work, binding interactions between biomolecules were analyzed by a technique that is based on electrically controllable DNA nanolevers. The technique was applied to virus-receptor interactions for the first time. As receptors, primarily peptides on DNA nanostructures and antibodies were utilized. The DNA nanostructures were integrated into the measurement technique and enabled the presentation of the peptides in a controllable geometrical order. The number of peptides could be varied to be compatible to the binding sites of the viral surface proteins. Influenza A virus served as a model system, on which the general measurability was demonstrated. Variations of the receptor peptide, the surface ligand density, the measurement temperature and the virus subtypes showed the sensitivity and applicability of the technology. Additionally, the immobilization of virus particles enabled the measurement of differences in oligovalent binding of DNA-peptide nanostructures to the viral proteins in their native environment. When the coronavirus pandemic broke out in 2020, work on binding interactions of a peptide from the hACE2 receptor and the spike protein of the SARS-CoV-2 virus revealed that oligovalent binding can be quantified in the switchSENSE technology. It could also be shown that small changes in the amino acid sequence of the spike protein resulted in complete loss of binding. Interactions of the peptide and inactivated virus material as well as pseudo virus particles could be measured. Additionally, the switchSENSE technology was utilized to rank six antibodies for their binding affinity towards the nucleocapsid protein of SARS-CoV-2 for the development of a rapid antigen test device. The technique was furthermore employed to show binding of a non-enveloped virus (adenovirus) and a virus-like particle (norovirus-like particle) to antibodies. Apart from binding interactions, the use of DNA origami levers with a length of around 50 nm enabled the switching of virus material. This proved that the technology is also able to size objects with a hydrodynamic diameter larger than 14 nm. A theoretical work on diffusion and reaction-limited binding interactions revealed that the technique and the chosen parameters enable the determination of binding rate constants in the reaction-limited regime. Overall, the applicability of the switchSENSE technique to virus-receptor binding interactions could be demonstrated on multiple examples. While there are challenges that remain, the setup enables the determination of affinities between viruses and receptors in their native environment. Especially the possibilities regarding the quantification of oligo- and multivalent binding interactions could be presented.}, language = {en} }