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The diploma thesis aimed to create a concept of a universal vehicle meeting the requirements for safe transportation of patients with risk of environmental contamination including the possibility of repeated use. The vehicle should meet the highest degree of public health protection, namely the Biological safety levels 4 (BSL 4). The questionnaire was used to gather the information about the current status of Biohazard teams, their material and technical equipment, and the way of transport security, including the possibility of rotating/substitution of the intervening staff. The questionnaire was distributed to the providers of health rescue services in individual regions and in the capital city of Prague. The results of the questionnaire survey formed the basis for processing the concept of the vehicle to ensure the transport of patients suspected of a highly dangerous contagion. The concept of the vehicle should serve as the basis for the real vehicle construction that will be able to provide protection at the level of BSL 4. It counts on a separate cabin for two drivers, a rear cabinet superstructure with a clean area for storing material and medical equipment, and rest facilities for the staff. At the rear, a separate insulating container consisting of an insulating box for patients, a decontamination area for the needs of the rotating staff, and a transfer box for medical supplies and devices. The proposed vehicle brings numerous improvements that meet BSL 4 protection and thus the highest level of the public health protection. The vehicle can carry up to seven patients at one intervention with the risk of environmental contamination. During the transport, it is possible for the staff to rotate continuously. The separate storage of medical supplies and devices ensures the use and degradation of only the necessary materials and devices. To cover the territory of the Czech Republic, the optimal number of six vehicles would be optimal.

<p dir="ltr">The recent global pandemic, COVID-19, has revealed to all the importance of understanding the complex relationship between viruses and hosts. Before COVID-19, I started my study of viral protein-host lipid interactions in the hemorrhagic fevers Ebola and Marburg viruses. These viruses contain a matrix protein that interacts with the plasma membrane to facilitate the formation of both authentic viruses and virus-like particles. My goal was to understand the limitations of their specific host lipid interactions. However, when the COVID-19 pandemic began, so to be our swift response in the development of a biosafety level 2 compatible model. This model can be used for studying severe acute respiratory distress syndrome 2 (SARS-CoV-2) assembly, egress, and entry. This model enabled exponentially greater access to more facilities to study the intricacies of SARS-CoV-2 assembly. With more access to studying the virus in a safe model, our goal is to push the understanding of viral assembly faster. I then began to take apart the individual pieces of the model and started to look at understanding the roles that they play independently. The membrane protein is the most abundant structural protein and I studied the specific lipid interactions of the soluble fraction of the protein. Physicians observed nucleocapsid protein mutations in the clinic with the increasing number of SARS-CoV-2 variants that are on the rise. The microscopy data collected can give us more insight into perhaps how the nucleocapsid protein induces the formation of filopodia structures at the plasma membrane. The envelope protein proved to be a challenge, but I determined a specific envelope and ceramide interaction in cells. The envelope protein was also causing the formation of microvesicles for an undefined function. I was able to determine the subcellular localization of the protein to the mitochondria. The localization to the mitochondria appears to induce depolarization of the mitochondria membrane action potential and induces the increase in mitochondria dysfunction signal, cytochrome c. Although the mitochondria were dysfunctional, there was no increase in apoptosis signal in the presence of the protein alone.</p>