OBJECTIVES: The development of External Quality Assessment Schemes (EQAS) for clinical flow cytometry (FCM) is challenging in the context of rare (immunological) diseases. Here, we introduce a novel EQAS monitoring the primary immunodeficiency Orientation Tube (PIDOT), developed by EuroFlow, in both a 'wet' and 'dry' format. This EQAS provides feedback on the quality of individual laboratories (i.e., accuracy, reproducibility and result interpretation), while eliminating the need for sample distribution. METHODS: In the wet format, marker staining intensities (MedFIs) within landmark cell populations in PIDOT analysis performed on locally collected healthy control (HC) samples, were compared to EQAS targets. In the dry format, participants analyzed centrally distributed PIDOT flow cytometry data (n=10). RESULTS: We report the results of six EQAS rounds across 20 laboratories in 11 countries. The wet format (212 HC samples) demonstrated consistent technical performance among laboratories (median %rCV on MedFIs=34.5 %; average failure rate 17.3 %) and showed improvement upon repeated participation. The dry format demonstrated effective proficiency of participants in cell count enumeration (range %rCVs 3.1-7.1 % for the major lymphoid subsets), and in identifying lymphoid abnormalities (79.3 % alignment with reference). CONCLUSIONS: The PIDOT-EQAS allows laboratories, adhering to the standardized EuroFlow approach, to monitor interlaboratory variations without the need for sample distribution, and provides them educational support to recognize rare clinically relevant immunophenotypic patterns of primary immunodeficiencies (PID). This EQAS contributes to quality improvement of PID diagnostics and can serve as an example for future flow cytometry EQAS in the context of rare diseases.

• MeSH
• Immunophenotyping * standards   methods   MeSH
• Humans MeSH
• Flow Cytometry * standards   methods   MeSH
• Quality Control MeSH
• Immunologic Deficiency Syndromes diagnosis   immunology   MeSH
• Quality Assurance, Health Care MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH

OBJECTIVE: Little is known about the association between subjectively experienced levels of diabetes distress (DD) and personality traits (PTs), even when levels of DD appear stable over time. This study aimed to use the Alternative Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition (DSM-5) Model for Personality Disorders (AMPD) to associate specific maladaptive PTs with experienced DD and to describe differences in the constellation of PTs between people with type 1 diabetes (PWT1D) and type 2 diabetes (PWT2D). METHODS: A total of 358 participants with diabetes mellitus (DM) (56.2% female, mean age 42.33 years, standard deviation (SD) = 14.33) were evaluated using the Diabetes Distress Scale (DDS) and the shortened 160-item version of the Personality Inventory for DSM-5 (PID-5). Psychometric properties of the DDS were evaluated first, then the association between DDS and PID-5 scores, and the differences between groups based on diabetes type and DD level, were analyzed. RESULTS: Strong associations were found between the PID-5 Negative Affectivity (NEF) domain and the emotional burden (β = 0.852, pHolm < 0.001) and regimen distress (β = 0.435, pHolm = 0.006) DDS subscale scores. PWT1D had a higher level of personality pathology than PWT2D, as did participants with elevated levels of DD across most domains and facets of PID-5. CONCLUSIONS: Our findings suggest that attention should be paid to the level of NEF among people with diabetes in relation to their emotional burden and perception of regimen distress. We recommend a distinction between people based on their diabetes type. Implications for clinical practice and interventions for DD perceived through the lens of the dimensional DSM-5 PT model are discussed.

• Publication type
• Journal Article MeSH

Vrozené poruchy imunitního systému (PID/IEIs) představují dynamicky se rozvíjející skupinu onemocnění, která je kromě zvýšené náchylnosti k infekcím charakterizována také výskytem autoimunitních, autoinflamatorních, alergických a maligních komplikací. V dnešní době bylo popsáno kolem 500 těchto onemocnění a jejich počet se neustále zvyšuje. Výsledkem dynamickému pokroku v diagnostice a terapii těchto onemocnění je rozvoj genetické diagnostiky, zavedení screeningového vyšetření PID/IEIs u novorozenců, stejně tak jako používání nových terapeutických přístupů včetně genové terapie. V České republice bylo na počátku roku 2024 zavedeno plošné screeningové vyšetření těžké kombinované imunodeficience (SCID). Rozvoj nových technologií umožňuje zlepšení genetické diagnostiky a získávání nových poznatků týkajících se patogeneze PID/IEIs, které lze využít k vývoji cílené terapie. Symptomatická léčba pacientů s PID/IEIs je kromě antimikrobiální profylaxe a imunoglobulinové substituční léčby obohacena o terapii tzv. malými molekulami a biologickou léčbou monoklonálními protilátkami namířenými proti buňkám nebo jejich produktům. V kauzální léčbě pacientů s PID/IEIs dochází ke zlepšení postupů v provádění transplantace hematopoetických krevních buněk (HSCT) a pokrokům na poli genové terapie. Budoucí vývoj v oblasti PID/IEIs bude směřovat k dalšímu zpřesnění jejich diagnostiky a rozvoji cílené terapie včetně personalizovaného přístupu k léčbě.

Congenital disorders of the immune system PIDs/IEIs) represent a dynamically developing group of diseases characterized not only by an increased susceptibility to infections but also by the occurrence of autoimmune, autoinflammatory, allergic, and malignant complications. Around 500 of these diseases have been described to date, and their number continues to grow. The result of dynamic progress in the diagnosis and therapy of these diseases is the progress of genetic diagnostics, the introduction of screening for PID/IEIs in newborns, as well as the use of new therapeutic approaches including gene therapy. Since the beginning of 2024, nationwide screening for severe combined immunodeficiency (SCID) has been introduced in the Czech Republic. Advances in genetic diagnostic technologies have allowed for more precise diagnostics and the acquisition of new data regarding the pathogenesis of PIDs/IEIs, which can be used to develop targeted therapies. Symptomatic treatment for patients with PIDs/IEIs, in addition to antimicrobial prophylaxis and immunoglobulin replacement therapy, has been enhanced with small molecules and biological therapy using monoclonal antibodies targeted against cytokines, cells, or their products. In the causal treatment of patients with PIDs/IEIs, there have been improvements in the procedures for hematopoietic stem cell transplantation (HSCT) and advances in the field of gene therapies. Future developments in the field of PID/IEIs will focus on more precise diagnostics, targeted therapies, and a personalized approach to treatment.

• MeSH
• Genetic Therapy MeSH
• Humans MeSH
• Antibodies, Monoclonal pharmacology   therapeutic use   MeSH
• Infant, Newborn MeSH
• Mass Screening MeSH
• Primary Immunodeficiency Diseases * diagnosis   therapy   MeSH
• Check Tag
• Humans MeSH
• Infant, Newborn MeSH
• Publication type
• Review MeSH

BACKGROUND: The safety, tolerability, and immunogenicity of hyaluronidase-facilitated subcutaneous immunoglobulin (fSCIG) 10% (dual-vial unit of human immunoglobulin 10% and recombinant human hyaluronidase [rHuPH20]) were assessed in children with primary immunodeficiency diseases (PIDs). METHODS: This phase 4, post-authorization, prospective, interventional, multicenter study (NCT03116347) conducted in the European Economic Area, enrolled patients aged 2 to < 18 years with a documented PID diagnosis who had received immunoglobulin therapy for ≥ 3 months before enrollment. New fSCIG 10% starters underwent fSCIG 10% dose ramp-up for ≤ 6 weeks (epoch 1) before receiving fSCIG 10% for ≤ 3 years (epoch 2); patients pretreated with fSCIG 10% entered epoch 2 directly. The primary outcome was the number and rate (per infusion) of all noninfectious treatment-related serious and severe adverse events (AEs). RESULTS: In total, 42 patients were enrolled and dosed (median [range] age: 11.5 [3-17] years; 81% male; 23 new starters; 19 pretreated). Overall, 49 related noninfectious, treatment-emergent AEs (TEAEs) were reported in 15 patients; most were mild in severity (87.8%). No treatment-related serious TEAEs were reported. Two TEAEs (infusion site pain and emotional distress) were reported as severe and treatment-related in a single new fSCIG 10% starter. The rate of local TEAEs was lower in pretreated patients (0.1 event/patient-year) versus new starters (1.3 events/patient-year). No patients tested positive for binding anti-rHuPH20 antibodies (titer of ≥ 1:160). CONCLUSIONS: No safety signals were identified, and the incidence of local AEs declined over the duration of fSCIG 10% treatment. This study supports fSCIG 10% long-term safety in children with PIDs. TRIAL REGISTRATION NUMBER (CLINICALTRIALS.GOV): NCT03116347.

• Publication type
• Journal Article MeSH

Acute respiratory distress syndrome (ARDS) is a disease that has a high reported mortality rate. The treatment for ARDS typically involves mechanical ventilation that is tailored to each patient's needs. A crucial aspect of this treatment is maintaining adequate oxygen saturation of haemoglobin by setting the fraction of inspired oxygen. This paper proposes a design method of robust proportional-integral-derivative (PID) controllers using a gas exchange model during ARDS. Several PID controllers were synthesized for different sub-operational ranges defined by measurable quantities of the mechanical ventilator and the patient using a mixed sensitivity H∞ approach. In simulations, the controller demonstrated high robustness to external changes and changes in the patient's condition, with saturation always above 88%. Although further validation of the controller is required, the results indicate that the presented robust control method has the potential to be clinically relevant.

Historik a novinář Peter Vronsky se snaží pochopit, jak nové poznatky o povaze člověka a sklonů k násilí zapadají do lidské historie sahající až do pravěku, což je podle něj klíčem k pochopení podstaty chování sériového vraha.

Primární imunodeficience (PID) je skupina geneticky podmíněných poruch imunitního systému, u které již bylo popsáno více než 300 kauzálních mutací. Negativním prognostickým faktorem pacientů s PID je tzv. dysregulace imunitního systému, projevující se autoreaktivními a autoinflamatorními komplikacemi. Zavedením sekvenování nové generace do klinického hodnocení je rychle rozšiřován rozsah monogenních poruch odpovědných za rozvoj PID. Značná část takto nalezených genetických variant ale není popsána v literatuře ani v dostupných databázích, a je proto bez předchozího testování nemožné posoudit jejich vliv na patogenezi a progresi onemocnění. Proto navrhujeme vyvinout a zavést do klinického hodnocení i metody podrobné imunofenotypizace leukocytů, doplněné o funkční hodnocení jejich reakcí na ex vivo stimuly. Souběžně budeme sekvenovat repertoár T a B buněčných receptorů pro identifikaci jejich restrikce, která se může podílet na projevech dysregulace. Souhrnné poznatky o změnách ve fenotypu, signalizaci a imunitním repertoáru pomůžou vybrat a v čase monitorovat vhodnou léčbu PID.; Primary Immunodeficiency Diseases (PIDD) are a group of disorders of the immune system, with over 300 causal mutations described to date. Immune dysregulation manifesting as autoimmunity and autoinflammation was described as a negative prognostic factor for patients with PIDD. Implementation of next generation sequencing into the clinical evaluation of affected individuals widens the range of identified monogenic errors rapidly. However, uncovering new variants previously undescribed in the literature and available databases makes impossible to assess their impact on the pathogenesis and progression of the disease. Therefore we propose to develop and implement methods for detailed immunophenotypic description of leukocytes, complemented with functional assessment of responses to ex vivo stimuli. In parallel, we will sequence a repertoire of T cell and B cell receptors in search for skewed patterns explaining deficiency and dysregulation. These insights into cellular, signaling and repertoire changes will enable us to select and monitor proper therapy in patients with PIDD.

• Keywords
• sekvenování nové generace, Next generation sequencing, primární imunodeficience, dysregulace, dysregulation, signalizace, imunofenotyp, immunophenotype, signaling, funkční testy, functional assays, repertoár receptorů lymfocytů, lymphocyte receptor repertoire, autoinflamatorní projevy, monogenní poruchy, Primary Immunodeficiency Diseases, autoinflammation, monogenic errors,

• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

The aim of this study was to determine the presence of Chlamydia trachomatis (ChT) and Neisseria gonorrhoae (NG) in the genital tract of women with ectopic pregnancy and to compare the positive results with patients' self-reported history of PID. Overall 40 women were eligible for the study. The samples for the ChT and NG Polymerase Chain-reaction (PCR) detection were obtained from the cervix, endometrium and fallopian tube. The results of testing for NG at all sites were negative as were the results from cervical testing for ChT. The prevalence of ChT in the upper genital tract was 12.5%. No statistically significant correlation was found between the positive cases and the previous signs of PID and laparoscopic findings. We found statistically significant relationship between signs of pelvic inflammation in a pacients' history and histopathological findings of tubal inflammation. 12.5% prevalence of ChT confirms the ascending genital infection. There was no association between the positive PCR result and clinical history of pelvic inflammation.IMPACT STATEMENTWhat is already known on this subject? Pelvic inflammatory disease, Chlamydia trachomatis and Neisseria gonorrhoae infections are the main risks for ectopic pregnancy. Clinical history of PID and perioperative adhesions may suggest prior upper genital infection.What do the results of this study add?Chlamydia trachomatis positive PCR result can be found in the upper genital tract without the positivity of cervical smear in women with ectopic pregnancy. Our study is unique in assessing the endometrial biopsy for the presence of Chlamydia trachomatis and Neisseria gonorrhoae.What are the implications of these findings for future clinical practice and/or future clinical research? There is no statistically significant association between positive PCR result and clinical history of PID or pelvic adhesions found during laparoscopy for tubal pregnancy. Therefore there is no need for the preventive antibiotic treatment in patients with these findings.

• MeSH
• Chlamydia trachomatis MeSH
• Chlamydia Infections * complications   diagnosis   epidemiology   MeSH
• Humans MeSH
• Pregnancy, Ectopic * diagnosis   epidemiology   MeSH
• Neisseria gonorrhoeae MeSH
• Pelvic Inflammatory Disease * complications   MeSH
• Polymerase Chain Reaction MeSH
• Pregnancy MeSH
• Inflammation complications   MeSH
• Check Tag
• Humans MeSH
• Pregnancy MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

• MeSH
• Autoantibodies * MeSH
• Common Variable Immunodeficiency * diagnosis   epidemiology   MeSH
• Immunologic Tests MeSH
• Humans MeSH
• Prevalence MeSH
• Check Tag
• Humans MeSH
• Publication type
• Letter MeSH
• Research Support, Non-U.S. Gov't MeSH

Primární imunodeficience (PID) jsou vzácná onemocnění imunitního systému, projevující se zejména zvýšenou náchylností k infekcím, jejich závažnějším průběhem, ale také projevy imunitní dysregulace, jako jsou lymfoproliferace či projevy autoimunity. Diagnostika PID zaznamenala v posledních letech obrovský rozvoj, umožněný zejména metodami sekvenování nové generace (NGS). Vliv nově identifikovaných genetických variant na imunitní systém však často není jednoznačný. V tomto projektu tedy navrhujeme rozšíření postupů pro genetickou diagnostiku pacientů s dosud neurčenými PID a následně rozvoj laboratorních metod umožňujících zkoumání funkčního vlivu nalezených variant. Práce bude navazovat na počáteční zkušenosti s pacienty s komplexními imunodeficity na řešitelském pracovišti. Přesná znalost genetické poruchy a jejího funkčního vlivu nám poté umožní navrhnout nové terapeutické postupy, cílené na konkrétní molekulární podstatu onemocnění pacientů. Tato specifická léčba může příznivě ovlivnit i závažné stavy imunitní dysregualce rezistentní k běžně používaným lékům.; Primary immunedeficiencies (PIDs) are rare diseases of the immune system, manifesting primarily as increased susceptibility to infections, their more severe course, but also immune dysregulations, such as lymphoproliferation or autoimmunity. The diagnostic of PID experienced a massive boom in recent years, enabled by the next-generation sequencing (NGS) methods. However, the impact of newly identified genetic variants on the immune system is frequently uncertain. In this project, we propose to expand the pipeline for genetic diagnosis of patients with unspecified PIDs and subsequent laboratory evaluation of identified variant functional impact. The project will build upon the applicant department’s previous experience in diagnosing and treating patients with complex immunodeficiencies. The knowledge of precise genetic variant and its functional impact will allow us to propose novel therapeutic approaches, targetted at the specific molecular cause of the disease. This specific therapy can be beneficial in severe immune dysregulation resistant to commonly used medication.

• MeSH
• Molecular Medicine MeSH
• Primary Immunodeficiency Diseases diagnosis   genetics   MeSH
• High-Throughput Nucleotide Sequencing MeSH

• Conspectus
• Patologie. Klinická medicína
• NML Fields
• alergologie a imunologie
• genetika, lékařská genetika
• molekulární biologie, molekulární medicína
• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR