Východiska: Brachyterapie (BRT) je nedílnou součástí radikální radioterapie (RT) nebo radiochemoterapie (RCHT) u pacientek, které nejsou vhodnými kandidátkami k operačnímu řešení. Jedná se většinou o pacientky s lokálně pokročilými karcinomy děložního hrdla. Cílem celého snažení při plánování BRT bylo, stále je a jistě nadále bude co možná nejlépe definovat anatomické hranice nádoru a vztah nádoru k rizikovým orgánům (organs at risk – OAR), a to za použití dostupných moderních zobrazovacích metod. Obrazem řízená adaptivní brachyterapie (image guided adaptive brachytherapy – IGABT) je v současné době nejmodernějším způsobem aplikace uterovaginální BRT. Adaptivní plánování umožňuje navyšování dávky z BRT do nově definovaných cílových objemů, a to podle míry rizika rekurence, které je dáno zejména mírou nádorové nálože. Tato dávková adaptace podle odpovědi na zevní RCHT je zásadní změnou praxe ve srovnání s konvenčním plánováním BRT založeným na dávkové preskripci do bodu A. Hlavní předností konceptu IGABT je, že dovoluje hodnocení individuální dávkové distribuce v cílových objemech a OAR, což následně vede ke zlepšenému dávkovému pokrytí cílových objemů za současného poklesu objemu, který je ozářen předepsanou dávkou ve srovnání s konvenčním 2D plánováním. Cíl: V tomto přehledovém článku předkládám ucelený aktuální pohled na tuto problematiku, zejména ve smyslu praktických doporučení týkajících se definování cílových objemů, použití různých druhů uterovaginálních aplikátorů, intraoperačních komplikací a možných projevů pozdní gastrointestinální, genitourinární a vaginální toxicity.

Brachytherapy (BT) is an integral part of radical radiotherapy (RT) or radiochemotherapy (RCT) in patients who are not suitable candidates for surgery. These are usually patients with locally advanced cervical cancer. The goal of all BT planning eff orts has been, still is, and certainly will continue to be, to defi ne the anatomical boundaries of the tumor and the relationship of the tumor to organs at risk (OARs) as best as possible, using available modern imaging techniques. Image guided adaptive brachytherapy (IGABT) is currently the most advanced method of uterovaginal BT. Adaptive planning allows dose escalation from BT to newly defi ned target volumes, according to the risk of recurrence, which is mainly determined by the level of tumor burden. This dose adaptation based on the response to external RCT is a major change in practice compared to conventional BT planning based on dose prescription to point A. The main advantage of the IGABT concept is that it allows the assessment of individual dose distributions in target volumes and OARs, which in turn leads to improved dose coverage of target volumes while decreasing the volume irradiated by the prescribed dose compared to conventional 2D planning. Purpose: In this review article, I provide a comprehensive up-to-date perspective on this issue, particularly in terms of practical recommendations regarding the defi nition of target volumes, the use of diff erent types of uterovaginal applicators, intraoperative complications, and potential manifestations of late gastrointestinal, genitourinary, and vaginal toxicity.

• Klíčová slova
• uterovaginální brachyterapie,
• MeSH
• brachyterapie metody   MeSH
• lidé MeSH
• nádory děložního čípku * radioterapie   MeSH
• radioterapie metody   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• přehledy MeSH

Diverse repertoires of hypervariable immunoglobulin receptors (TCR and BCR) recognize antigens in the adaptive immune system. The development of immunoglobulin receptor repertoire sequencing methods makes it possible to perform repertoire-wide disease association studies of antigen receptor sequences. We developed a statistical framework for associating receptors to disease from only a small cohort of patients, with no need for a control cohort. Our method successfully identifies previously validated Cytomegalovirus and type one diabetes responsive TCR[Formula: see text] sequences .

• MeSH
• adaptivní imunita genetika   MeSH
• Cytomegalovirus imunologie   MeSH
• diabetes mellitus genetika   imunologie   MeSH
• genetická variace imunologie   MeSH
• hypervariabilní oblasti genetika   MeSH
• lidé MeSH
• receptory antigenů B-buněk genetika   MeSH
• receptory antigenů T-buněk genetika   imunologie   MeSH
• receptory antigenů genetika   imunologie   MeSH
• receptory imunologické genetika   imunologie   MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The design of smart surfaces with externally triggerable water/oil wettability and adhesion represents one of the most up-to-date challenges in the field of material science. In this work, the intelligent surface with electrically triggerable wettability and water/oil adhesion is presented. As a basic material background exhibiting electric field (EF) sensitivity, the piezo-responsive polymethylmethacrylate/polyvinylidenefluoride polymer fibers were used. To expand the available range of water/oil contact angles (CAs) and adhesion, the fibers were grafted with hydrophilic or hydrophobic functional groups using diazonium chemistry. The fiber functionality was evaluated using the static CA and wettability hysteresis measurements (increasing/decreasing drop volume and tilting angles), drops adhesion/repellence and graphite self-cleaning test performed with and without the application of EF. It was found that the proposed method enables tuning the surface wettability in the superhydrophobic/superoleophobic-hydrophilic/oleophilic range and changing of surface properties from low adhesive to high adhesive for water and oil. More convincing results were achieved in the case of fiber surface modification by ADT-C8F17, which may result from a rearrangement of the grated -C6H4C8F17 functional group under the application of EF triggering. Moreover, the triggering which can be performed in the extremely fast way (the surface responds to the EF switching on/off in seconds) was found to be fully reversible. Finally, the additional tests indicate the satisfactory stability of created fiber-based coating against the mechanical treatment.

• MeSH
• adhezivita MeSH
• chytré materiály MeSH
• fluoridy MeSH
• hydrofobní a hydrofilní interakce MeSH
• polymethylmethakrylát MeSH
• smáčivost MeSH
• testování materiálů * MeSH
• Publikační typ
• práce podpořená grantem MeSH

Background: The Human Cell Differentiation Molecules (HCDM) organizes Human Leukocyte Differentiation Antigen (HLDA) workshops to test and name clusters of antibodies that react with a specific antigen. These cluster of differentiation (CD) markers have provided the scientific community with validated antibody clones, consistent naming of targets and reproducible identification of leukocyte subsets. Still, quantitative CD marker expression profiles and benchmarking of reagents at the single-cell level are currently lacking. Objective: To develop a flow cytometric procedure for quantitative expression profiling of surface antigens on blood leukocyte subsets that is standardized across multiple research laboratories. Methods: A high content framework to evaluate the titration and reactivity of Phycoerythrin (PE)-conjugated monoclonal antibodies (mAbs) was created. Two flow cytometry panels were designed: an innate cell tube for granulocytes, dendritic cells, monocytes, NK cells and innate lymphoid cells (12-color) and an adaptive lymphocyte tube for naive and memory B and T cells, including TCRγδ+, regulatory-T and follicular helper T cells (11-color). The potential of these 2 panels was demonstrated via expression profiling of selected CD markers detected by PE-conjugated antibodies and evaluated using 561 nm excitation. Results: Using automated data annotation and dried backbone reagents, we reached a robust workflow amenable to processing hundreds of measurements in each experiment in a 96-well plate format. The immunophenotyping panels enabled discrimination of 27 leukocyte subsets and quantitative detection of the expression of PE-conjugated CD markers of interest that could quantify protein expression above 400 units of antibody binding capacity. Expression profiling of 4 selected CD markers (CD11b, CD31, CD38, CD40) showed high reproducibility across centers, as well as the capacity to benchmark unique clones directed toward the same CD3 antigen. Conclusion: We optimized a procedure for quantitative expression profiling of surface antigens on blood leukocyte subsets. The workflow, bioinformatics pipeline and optimized flow panels enable the following: 1) mapping the expression patterns of HLDA-approved mAb clones to CD markers; 2) benchmarking new antibody clones to established CD markers; 3) defining new clusters of differentiation in future HLDA workshops.

• MeSH
• antigeny povrchové * metabolismus   MeSH
• buňky NK metabolismus   MeSH
• CD antigeny metabolismus   MeSH
• leukocyty MeSH
• lidé MeSH
• monoklonální protilátky MeSH
• přirozená imunita * MeSH
• průběh práce MeSH
• průtoková cytometrie metody   MeSH
• referenční standardy MeSH
• reprodukovatelnost výsledků MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Adjuvanticity, which is the ability of neoplastic cells to deliver danger signals, is critical for the host immune system to mount spontaneous and therapy-driven anticancer immune responses. One of such signals, i.e., the exposure of calreticulin (CALR) on the membrane of malignant cells experiencing endoplasmic reticulum (ER) stress, is well known for its role in the activation of immune responses to dying cancer cells. However, the potential impact of CALR on the immune contexture of primary and metastatic high-grade serous carcinomas (HGSCs) and its prognostic value for patients with HGSC remains unclear. METHOD: We harnessed a retrospective cohort of primary (no = 152) and metastatic (no = 74) tumor samples from HGSC patients to investigate the CALR expression in relation with prognosis and function orientation of the tumor microenvironment. IHC data were complemented with transcriptomic and functional studies on second prospective cohort of freshly resected HGSC samples. In silico analysis of publicly available RNA expression data from 302 HGSC samples was used as a confirmatory approach. RESULTS: We demonstrate that CALR exposure on the surface of primary and metastatic HGSC cells is driven by a chemotherapy-independent ER stress response and culminates with the establishment of a local immune contexture characterized by TH1 polarization and cytotoxic activity that enables superior clinical benefits. CONCLUSIONS: Our data indicate that CALR levels in primary and metastatic HGSC samples have robust prognostic value linked to the activation of clinically-relevant innate and adaptive anticancer immune responses.

• MeSH
• dospělí MeSH
• kalretikulin imunologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové mikroprostředí genetika   imunologie   MeSH
• nádory vaječníků genetika   imunologie   MeSH
• prognóza MeSH
• sekvenování transkriptomu MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• stres endoplazmatického retikula MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, U.S. Gov't, Non-P.H.S. MeSH

Cíl práce: Molekula CD154 (CD40L) je transmembránový glykoprotein přítomný především v aktivovaných lymfocytech T CD4+. Jeho receptorem je molekula CD40, která je v membráně lymfocytů B, ale též jiných buněk. Interakce CD154-CD40 je podmínkou optimálního rozvoje adaptivní imunitní reakce, jejím důsledkem je však i ovlivnění zánětlivého procesu. Porucha funkce CD154 je patognomická pro závažnou primární imunodeficienci – syndrom hyperimunoglobulinémie M. Odhalení abnormality CD154 je základem pro stanovení diagnózy tohoto syndromu. Materiál a metodika: Vypracovali jsme mikrometodu, která umožňuje průkaz CD154 na lymfocytech aktivovaných in vitro v plné krvi a srovnali ji s metodou vyšetření CD154 na izolovaných lymfocytech. Periferní heparinizovaná krev je inkubována 4 hodiny s aktivačními činidly (phorbol-myristat-acetat, ionomycin), inkubována s monoklonálními protilátkami a vyšetřena na průtokovém cytometru. Vzhledem k tomu, že v průběhu stimulace dochází ke ztrátě molekul CD4 v plazmatické membráně, jsou CD4+ lymfocyty identifikovány jako CD5+/CD8. Jejich aktivace je sledována expresí CD69. K simultánnímu hodnocení přítomnosti CD154 se používá tříbarevná průtoková cytometrie. Výsledky: Metodika byla aplikována u deseti zdravých dárců krve. K provedení testu stačí 0,5 ml heparinizované krve. Optimální doba aktivace, která umožňuje detegovatelnou expresi CD154 na lymfocytech T, jsou 4 hodiny. Zjistili jsme, že v průběhu této aktivace dochází ke snížení množství CD4 molekul na lymfocytech T. Hodnoty exprese CD154 jsou plně srovnatelné při použití naší metody s plnou krví i metody s izolovanými leukocyty. Závěr: Naše metodika k průkazu CD154 na lymfocytech aktivovaných v plné krvi je časově nenáročná, vyžaduje malé množství (0,5 ml) krve a lze ji doporučit při vstupním vyšetření dětí s podezřením na syndrom hyperimunoglobulinémie M. Umožňuje také záchyt přenašeček XHIGM.

Objective: CD154 (also called CD40L) is a transmembrane glycoprotein predominantly expressed on the surface membrane of activated CD4+ T cells. Its receptor CD40 is present on all B cells, but also on other cells. The interaction CD154-CD40 is necessary for the optimal development of the adaptive immune response and also has consequences for the modulation of the inflammatory response. A defect in the expression of CD154 is pathognomonic of congenital immunodeficiency called X-linked Hyper-IgM syndrome (XHIGM). To detect the abnormality of CD154 is essential for making the diagnosis of XHIGM. Material and methods: We worked out a microtest for the detection of CD154 expression on in vitro activated CD4+ T cells in whole blood and compared it with that on isolated cells from peripheral blood. Heparinized peripheral blood was activated with phorbol 12-myristate 13-acetate and ionomycin for 4 hours, labeled with monoclonal antibodies and analyzed by flow cytometry. Considering that the CD4 marker on the plasma membrane surface decreases during the activation, CD4+ T cells are mostly recognized as CD5+/CD8- cells. Their activation is monitored based on the expression of CD69. Three-?-colour immunofluorescence staining was used for simultaneous detection of CD154. Results: Ten blood donors were tested. As little as 0.5 ml of heparinized whole blood is needed to complete the test. Optimal time for activation and detection of CD154 on T lymphocytes is 4 hours. We found that the number of CD4 molecules on the surface of T cells decreases during the activation. The expression of CD154 in our whole blood microtest is fully comparable with that in the test on isolated leukocytes. Conclusion: The presented microtest for the detection of CD154 on activated lymphocytes in whole blood is fast and blood saving, since as little as 0.5 ml of blood is needed to complete it. It can be recommended as the initial test for suspected hyper-IgM syndrome in children. We demonstrate that this screening method can help to detect also carriers of XHIGM.

• Klíčová slova
• syndrom hyperimunoglobulinémie M, CD40,
• MeSH
• aktivace lymfocytů imunologie   MeSH
• CD4-pozitivní T-lymfocyty imunologie   klasifikace   MeSH
• fluorescenční protilátková technika MeSH
• imunodeficience s hyper-IgM, typ 1 diagnóza   MeSH
• imunofenotypizace metody   MeSH
• lidé MeSH
• ligand CD40 analýza   MeSH
• lymfocyty imunologie   klasifikace   MeSH
• průtoková cytometrie MeSH
• Check Tag
• lidé MeSH

BACKGROUND: Allogeneic hematopoietic stem cell transplant (allo-HSCT) recipients are at high risk of complications associated with COVID-19 infection due to dysfunction of their immune system. Vaccination can protect from the adverse consequences of COVID-19. However, studies on the efficacy of COVID-19 vaccines in HSCT recipients with insufficient post-HSCT immune reconstitution are still scarce. In our study, we determined how immunosuppressive medication and the reconstitution of the cellular immune system influenced T cell responses specific for the surface glycoprotein of SARS-CoV-2 virus (S antigen) after two doses of mRNA vaccine against COVID-19 in patients with myeloid malignancies treated with HSCT. METHODS: Vaccination outcomes were followed in 18 (allo-HSCT) recipients and 8 healthy volunteers. The IgG antibodies against SARS-CoV-2 spike (S) and nucleocapsid (NCP) protein were determined in ELISA and S-specific T cells were detected using a sensitive ELISPOT-IFNγ based on in vitro expansion and restimulation of T cells in pre- and post-vaccination blood samples. Multiparametric flow cytometry analysis of peripheral blood leukocyte differentiation markers was employed for determination of reconstitution of the main subpopulations of T cells and NK cells at month 6 after HSCT. RESULTS: S- specific IgG antibody response detected in 72% of the patients was lower than in healthy vaccinees (100%). Vaccine-induced T-cell responses to S1 or S2 antigen were significantly reduced in HSCT recipients, which were treated with corticosteroids in dose 5 mg of prednisone- equivalents or higher during the vaccination period or in preceeding 100 days in comparison with recipients un-affected with corticosteroids. A significant positive correlation was found between the level of anti-SARS-Cov-2 spike protein IgG antibodies and the number of functional S antigen-specific T cells. Further analysis also showed that the specific response to vaccination was significantly influenced by the interval between administration of vaccine and transplantation. Vaccination outcomes were not related to age, sex, type of mRNA vaccine used, basic diagnosis, HLA match between HSC donor and recipient, and blood counts of lymphocytes, neutrophils, and monocytes at the time of vaccination. Multiparametric flow cytometry analysis of peripheral blood leukocyte differentiation markers showed that good humoral and cellular S-specific immune responses induced by vaccination were associated with well-reconstituted CD4+ T cells, mainly CD4+ effector memory subpopulation at six months after HSCT. CONCLUSIONS: The results showed that both humoral and cellular adaptive immune responses of HSCT recipients to the SARS-CoV-2 vaccine were significantly suppressed by corticosteroid therapy. Specific response to the vaccine was significantly affected by the length of the interval between HSCT and vaccination. Vaccination as early as 5 months after HSCT can lead to a good response. Immune response to the vaccine is not related to age, gender, HLA match between HSC donor and recipient, or type of myeloid malignancy. Vaccine efficacy was dependent on well-reconstituted CD4+ T cells, at six months after HSCT.

• MeSH
• COVID-19 * prevence a kontrola   MeSH
• imunita MeSH
• imunoglobulin G MeSH
• imunosupresivní léčba MeSH
• lidé MeSH
• mRNA vakcíny MeSH
• nádory * MeSH
• SARS-CoV-2 MeSH
• transplantace hematopoetických kmenových buněk * škodlivé účinky   MeSH
• vakcíny proti COVID-19 MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Janis Kuby’s groundbreaking introduction to immunology was the first textbook for the course actually written to be a textbook. Like no other text, it combined an experimental emphasis with extensive pedagogical features to help students grasp basic concepts.Now in a thoroughly updated new edition, Kuby Immunology remains the only undergraduate introduction to immunology written by teachers of the course. In the Kuby tradition, authors Judy Owen, Jenni Punt, and Sharon Stranford present the most current concepts in an experimental context, conveying the excitement of scientific discovery, and highlight important advances, but do so with the focus on the big picture of the study of immune response, enhanced by unsurpassed pedagogical support for the first-time learner.

• MeSH
• alergologie a imunologie MeSH
• imunitní systém MeSH
• Publikační typ
• učebnice MeSH

• Konspekt
• Patologie. Klinická medicína
• Učební osnovy. Vyučovací předměty. Učebnice
• NLK Obory
• alergologie a imunologie

BACKGROUND: Ageing commonly disrupts the balance control and compensatory postural responses that contribute to maintaining balance and preventing falls during perturbation of posture. Improvement of compensatory postural responses during walking is one of the main goals in fall prevention programs which often include treadmill walking training. However, during treadmill walking, there is a sensory (visualsomatosensory and vestibular-somatosensory) conflict that can evoke aftereffects of self-motion sensation and could alter postural stability after training. RESEARCH QUESTION: The aim of this study was to compare the effect of overground and treadmill walking on postural stability in healthy young and elderly subjects. METHODS: Postural responses of 31 Young and 19 healthy Elderly before and after overground and treadmill walking were assessed by a force platform in four stance conditions: firm and foam surface with eyes open and eyes closed. RESULTS: In Elderly group, velocity parameters significantly increased after treadmill walking but not after overground walking. This increase was found particularly in the conditions with eyes open in both types of surfaces (firm, foam). The velocity parameters values (expect Vx) were significantly increased in Elderly compared to Young almost in all four conditions after treadmill and overground walking. SIGNIFICANCE: Our study suggests that Elderly become more unstable after treadmill walking and have greater difficulties to adapt to new balance circumstances caused by sensory conflict associated with treadmill walking. It seems that during treadmill walking and subsequent stance, vision is the major factor contributing to posture stabilization. Thus, the suitability of treadmill walking as a part of training programs for elderly adults with higher fall risk should be seriously considered.

• MeSH
• chůze * MeSH
• čití, cítění MeSH
• fyziologická adaptace MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• posturální rovnováha * MeSH
• senioři MeSH
• stárnutí fyziologie   MeSH
• úrazy pádem prevence a kontrola   MeSH
• zátěžový test MeSH
• zrak MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Low iron bioavailability is a common feature of ocean surface water and therefore micro-algae developed original strategies to optimize iron uptake and metabolism. The marine picoeukaryotic green alga Ostreococcus tauri is a very good model for studying physiological and genetic aspects of the adaptation of the green algal lineage to the marine environment: it has a very compact genome, is easy to culture in laboratory conditions, and can be genetically manipulated by efficient homologous recombination. In this study, we aimed at characterizing the mechanisms of iron assimilation in O. tauri by combining genetics and physiological tools. Specifically, we wanted to identify and functionally characterize groups of genes displaying tightly orchestrated temporal expression patterns following the exposure of cells to iron deprivation and day/night cycles, and to highlight unique features of iron metabolism in O. tauri, as compared to the freshwater model alga Chalamydomonas reinhardtii. RESULTS: We used RNA sequencing to investigated the transcriptional responses to iron limitation in O. tauri and found that most of the genes involved in iron uptake and metabolism in O. tauri are regulated by day/night cycles, regardless of iron status. O. tauri lacks the classical components of a reductive iron uptake system, and has no obvious iron regulon. Iron uptake appears to be copper-independent, but is regulated by zinc. Conversely, iron deprivation resulted in the transcriptional activation of numerous genes encoding zinc-containing regulation factors. Iron uptake is likely mediated by a ZIP-family protein (Ot-Irt1) and by a new Fea1-related protein (Ot-Fea1) containing duplicated Fea1 domains. The adaptation of cells to iron limitation involved an iron-sparing response tightly coordinated with diurnal cycles to optimize cell functions and synchronize these functions with the day/night redistribution of iron orchestrated by ferritin, and a stress response based on the induction of thioredoxin-like proteins, of peroxiredoxin and of tesmin-like methallothionein rather than ascorbate. We briefly surveyed the metabolic remodeling resulting from iron deprivation. CONCLUSIONS: The mechanisms of iron uptake and utilization by O. tauri differ fundamentally from those described in C. reinhardtii. We propose this species as a new model for investigation of iron metabolism in marine microalgae.

• MeSH
• biologická adaptace MeSH
• Chlorophyta klasifikace   genetika   metabolismus   MeSH
• Eukaryota genetika   metabolismus   MeSH
• fotoperioda MeSH
• fylogeneze MeSH
• fytoplankton genetika   metabolismus   MeSH
• fyziologický stres MeSH
• homeostáza MeSH
• měď metabolismus   MeSH
• oxidace-redukce MeSH
• regulace genové exprese účinky záření   MeSH
• rostlinné proteiny genetika   metabolismus   MeSH
• shluková analýza MeSH
• signální transdukce MeSH
• sloučeniny železa metabolismus   MeSH
• stanovení celkové genové exprese MeSH
• transkriptom MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• železo metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH