Key Features Relevant to Select Antigens and TCR From the MHC-Mismatched Repertoire to Treat Cancer

Adoptive transfer of T cells transgenic for tumor-reactive T-cell receptors (TCR) is an attractive immunotherapeutic approach. However, clinical translation is so far limited due to challenges in the identification of suitable target antigens as well as TCRs that are concurrent safe and efficient. Definition of key characteristics relevant for effective and specific tumor rejection is essential to improve current TCR-based adoptive T-cell immunotherapies. We here characterized in-depth two TCRs derived from the human leukocyte antigen (HLA)-mismatched allogeneic repertoire targeting two different myeloperoxidase (MPO)-derived peptides presented by the same HLA-restriction element side by side comprising state of the art biochemical and cellular in vitro, in vivo, and in silico experiments. In vitro experiments reveal comparable functional avidities, off-rates, and cytotoxic activities for both TCRs. However, we observed differences especially with respect to cytokine secretion and cross-reactivity as well as in vivo activity. Biochemical and in silico analyses demonstrate different binding qualities of MPO-peptides to the HLA-complex determining TCR qualities. We conclude from our biochemical and in silico analyses of peptide-HLA-binding that rigid and high-affinity binding of peptides is one of the most important factors for isolation of TCRs with high specificity and tumor rejection capacity from the MHC-mismatched repertoire. Based on our results, we developed a workflow for selection of such TCRs with high potency and safety profile suitable for clinical translation

Задержки в диагностике и лечении у больных туберкулезом легких в период пандемии COVID-19 в г. Душанбе, Таджикистан, 2022

The objective: to study delays in provision of medical care to pulmonary tuberculosis patients above 15 years old with laboratory confirmation of the diagnosis before and during the COVID-19 pandemic in Dushanbe, Tajikistan.Subjects and Methods. A retrospective cohort study was conducted among new pulmonary tuberculosis patients above 15 years old with laboratory confirmation of the diagnosis registered by TB Control Centers in Dushanbe in 2019–2021. Information was collected from medical records, medical databases, and patient questionnaires. Two types of delays were detected during the provision of medical care: delays related to the patient (defined as >14 days between onset of tuberculosis symptoms and the first visit to a health unit); delays related to health system (defined as >3 days from the first visit to TB unit to the initiation of tuberculosis treatment).Results. Of the 472 patients enrolled in the study, 276 were registered during the COVID-19 pandemic (2020–2021) and 196 patients were registered before the pandemic (2019). Frequency of delays was similar (83% vs 82%, p=0.8) during and before the COVID-19 pandemic. The frequency of delay related to health system was statistically significantly lower during the pandemic versus the period before the pandemic (34% vs 44%, p=0.023). The median duration of delays associated to the patient before the pandemic (60 days, IQR: 15–541) and during the pandemic (60 days, IQR: 15–360, р=0.6) was also similar as well as delays related to health system (before the pandemic – 7 days, IQR: 4–336) and during the pandemic (7 days, IQR: 4–225, р=0.6).Conclusion. Contrary to the expected result, no significant difference in delays related to the patient before and during the pandemic was found, and the frequency of delay related to health system during the pandemic significantly reduced compared to the period before the pandemic.Цель исследования: изучить задержки при оказании медицинской помощи больным старше 15 лет с лабораторно подтвержденным туберкулезом легких до и в период пандемии CОVID-19 в г. Душанбе Республики Таджикистан.Материалы и методы. Проведено ретроспективное когортное исследование среди впервые выявленных лабораторно подтвержденных больных старше 15 лет с легочным туберкулезом, зарегистрированных в 2019–2021гг. в противотуберкулёзном центре г. Душанбе. Сведения получали из медицинских документов, медицинских баз данных и анкетирования пациентов. У больных при оказании медицинской помощи выявлялись задержки двух видов: задержка со стороны пациента (определялась как >14 дней между появлением симптомов туберкулеза и первым посещением медицинского учреждения); задержка со стороны системы здравоохранения (определялась как >3 дней от первого посещения противотуберкулезного учреждения до начала лечения туберкулеза).Результаты. Из 472 пациентов, включенных в исследование, 276 чел. были зарегистрированы во время пандемии COVID-19 (2020–2021 гг.) и 196 – до нее (2019 г.). Частота задержки со стороны пациентов была одинакова (83% против 82%, р=0,8) во время и до COVID-19. Частота задержки со стороны системы здравоохранения была статистически значимо реже в период пандемии, чем до неё (34% против 44%, p=0,023). Продолжительность задержек со стороны пациентов по медиане до пандемии (60 дней, IQR: 15–541) и во время (60 дней, IQR: 15–360, р=0,6) были сопоставимы также, как и задержки со стороны системы здравоохранения (до пандемии – 7 дней, IQR: 4–336) и во время (7 дней, IQR: 4–225), р=0,6.Заключение. Вопреки ожидаемому результату, существенной разницы в задержках со стороны пациентов до- и во время пандемии не установлено, а частота задержки со стороны системы здравоохранения во время пандемии значимо уменьшилась в сравнении с периодом до нее

Positive and negative effects of antipsychotic medication: an international online survey of 832 recipients

Background: Antipsychotic medication is currently the treatment of choice for psychosis, but few studies directly survey the first hand experience of recipients. Objectives: To ascertain the experiences and opinions of users of an international sample of antipsychotic drugs regarding positive and negative effects. Method: An online direct-to-consumer questionnaire was completed by 832 users of antipsychotics, from 30 countries – predominantly USA, UK and Australia. This is the largest such sample to date. Results: Over half (56%) thought the drugs reduced the problems they were prescribed for, but 27% thought they made them worse. Slightly less people found the drugs generally ‘Helpful’ (41%) than found them ‘Unhelpful’ (43%). While 35% reported that their ‘quality of life’ was ‘improved’, 54% reported that it was made ‘worse’. The average number of adverse effects reported was 11, with an average of five at the ‘severe’ level. Fourteen effects were reported by 57% or more participants, most commonly: ‘Drowsiness, feeling tired, sedation’ (92%), ‘Loss of motivation’ (86%), ‘Slowed thoughts’ (86%), and ‘Emotional numbing’ (85%). Suicidality was reported to be a side effect by 58%. Older people reported particularly poor outcomes and high levels of adverse effects. Duration of treatment was unrelated to positive outcomes but significantly related to negative outcomes. Most respondents (70%) had tried to stop taking the drugs. The most common reasons people wanted to stop were the side effects (64%) and worries about long-term physical health (52%). Most (70%) did not recall being told anything at all about side effects. Conclusions Clinical implications are discussed, with a particular focus on the principles of informed consent, and involving patients in decision making about their own lives

Inclusion of maintenance energy improves the intracellular flux predictions of CHO

Chinese hamster ovary (CHO) cells are the leading platform for the production of biopharmaceuticals with human-like glycosylation. The standard practice for cell line generation relies on trial and error approaches such as adaptive evolution and high-throughput screening, which typically take several months. Metabolic modeling could aid in designing better producer cell lines and thus shorten development times. The genome-scale metabolic model (GSMM) of CHO can accurately predict growth rates. However, in order to predict rational engineering strategies it also needs to accurately predict intracellular fluxes. In this work we evaluated the agreement between the fluxes predicted by parsimonious flux balance analysis (pFBA) using the CHO GSMM and a wide range of 13C metabolic flux data from literature. While glycolytic fluxes were predicted relatively well, the fluxes of tricarboxylic acid (TCA) cycle were vastly underestimated due to too low energy demand. Inclusion of computationally estimated maintenance energy significantly improved the overall accuracy of intracellular flux predictions. Maintenance energy was therefore determined experimentally by running continuous cultures at different growth rates and evaluating their respective energy consumption. The experimentally and computationally determined maintenance energy were in good agreement. Additionally, we compared alternative objective functions (minimization of uptake rates of seven nonessential metabolites) to the biomass objective. While the predictions of the uptake rates were quite inaccurate for most objectives, the predictions of the intracellular fluxes were comparable to the biomass objective function.COMET center acib: Next Generation Bioproduction, which is funded by BMK, BMDW, SFG, Standortagentur Tirol, Government of Lower Austria and Vienna Business Agency in the framework of COMET - Competence Centers for Excellent Technologies. The COMET-Funding Program is managed by the Austrian Research Promotion Agency FFG; D.S., J.S., M.W., M.H., D. E.R. This work has also been supported by the PhD program BioToP of the Austrian Science Fund (FWF Project W1224)info:eu-repo/semantics/publishedVersio