Infection of 3D Brain Organoids with Human Pathogenic Viruses Under Biosafety Level-3 Conditions with Subsequent Inactivation to Study Viral Replication, Pathomechanisms, and Other Viral Infection-Mediated Effects

Viral infectious diseases may cause neurological symptoms primarily in two nonexclusive ways. Infection may lead to an excessive inflammatory response that damages the neuronal system, which is referred to as immunopathology, or the pathogen is able to infect brain cells, such as neurons or nonneuronal glial cells, like oligodendrocytes, microglia, and astrocytes. Viruses that target and infect cells of the CNS, in general, are called neurotropic. 3D brain organoids provide favorable conditions to study target cells and induced pathomechanisms associated with such viral neurotropism. In the context of highly pathogenic viruses, strict safety precautions (safety level-3 laboratory) must be taken if infectious laboratory strains or strains from clinical samples are to be used for infection experiments. Likewise, safe inactivation protocols must be used for subsequent analysis. This chapter will discuss appropriate protocols, focusing on methodological aspects for each of these steps, and discuss advantages and disadvantages when working with 3D brain organoids while handling biosafety level-3 pathogens based on our work with severe acute respiratory virus type 2 (SARS-CoV-2). Copyright © 2023, Springer Science+Business Media, LLC, part of Springer Nature.

Updated results of the COVID-19 in MS global data sharing initiative validate consistent associations of anti-CD20 and other reported risk factors with severe COVID-19 outcomes

Background: As the COVID-19 pandemic continues, evidencebased clinical guidance for managing the care of people with multiple sclerosis (MS) is an ongoing concern. In recent months, data from cohorts of people with MS has indicated that certain demographic and clinical characteristics, including use of some disease- modifying therapies (DMTs), leads to worse outcomes from SARS-CoV-2 infection. The COVID-19 in MS global data sharing initiative, which now includes over 4,500 confirmed COVID- 19 cases in people with MS, gives the opportunity to corroborate previous findings with greater certainty. Methods: Clinician-reported data from 32 countries were aggregated into a dataset of 5,543 patients who had suspected or confirmed COVID-19. Demographic and clinical covariates were queried, alongside COVID-19 clinical severity outcomes. These outcomes (hospitalisation, admission to ICU, requiring artificial ventilation, and death) were assessed in patients with suspected/ confirmed COVID-19 using multilevel mixed-effects logistic regression. All models were corrected for age, sex, EDSS, and MS type. DMTs were individually compared to glatiramer acetate (GA), as well as to pooled other DMTs and natalizumab. Results: Of 5,543 patients in the clinician-reported dataset, 909 with suspected and 4,634 with confirmed COVID-19 were included in the analysis. Previous demographic findings were confirmed: male sex, older age, progressive MS, and higher disability were associated with worse outcomes from SARS-CoV-2 infection. Use of anti-CD20 DMTs (ocrelizumab and rituximab) was associated with worse COVID-19 outcomes. Compared to GA, ocrelizumab and rituximab were associated with increased risk of hospitalisation (aOR=1.61(95%CI=1.06-2.43);aOR=2.42(95%CI=1.54-3.81) and ICU admission (aOR=3.13(95%CI=1.22-8.00);aOR=4.46 (95%CI=1.64-12.09)). Rituximab was associated with increased risk of artificial ventilation (aOR=3.57(95%CI=1.38-9.20));ocrelizumab showed a positive trend (aOR=1.86(95%CI=0.76-4.55). Rituximab showed a positive trend with increased risk of death (aOR=2.74(95%CI=0.68-11.09). Associations persisted on restriction to confirmed COVID-19 cases. Conclusions: Analysing the largest international real world dataset of people with MS who have suspected or confirmed COVID- 19 confirms previous findings that male sex, older age, progressive MS, higher disability, the use of anti-CD20 medication (ocrelizumab and rituximab) are associated with worse COVID-19 outcomes.

Family physicians in training: Which information sources do they use to address the covid-19 pandemic?

Background In the context of (the corona pandemic induced) challenges, which physicians in further training (DIT) faced in the first phase of the corona pandemic, the Competence Centre for Further Training in General Practice Saxony (KWASa) developed a survey on the handling of information on the corona virus perceived at that time. The survey focused on the evaluation of formal and informal information provision on prevention in primary care, diagnostics and therapy of SARS-CoV-2. In addition, the DITs rated the quality of information and institutional support. Methods An online survey was conducted during 05/05/2020 to 04/06/2020 among the DITs registered in the KWASa since 2018. The questionnaire consisted of standardized elements, which were analyzed descriptively, and open items with free-text answers, which were analyzed according to the principle of qualitative content analysis. Results The results show that the DITs received little external support and mostly had to resort to their own information research. Only 26 % of the respondents stated that they had received active support in the form of information. The information carriers were assessed in terms of active dissemination of information and the quality of information. The Robert Koch Institute (RKI) and specialist journals were rated as most helpful. The results of the free-text answers show the high demand for information in such a pandemic situation. The lack of information on topics such as prevention, education on the effects on everyday life, study com-parisons and training based on case studies was criticized several times. A corresponding need for support was articulated. Conclusions Based on this study, the support needs of DITs in crisis situations such as the corona pandemic were identified. The results of the survey can be used to develop suitable further training programs for DITs.