BACKGROUND: The current generation of neonatal ventilators enables periodic storage of set, measured, and calculated ventilatory parameters. DESIGN: Retrospective observational study. OBJECTIVES: To evaluate and identify the ventilatory, demographic, and clinical pre-extubation variables that are significant for estimating extubation readiness. METHODS: Eligible subjects included premature infants <33 weeks of gestation weaned from mechanical ventilation (MV) lasting >24 h. A total of 16 relevant ventilator variables, each calculated from 288 data points over 24 h, together with eight demographic and three clinical pre-extubation variables, were used to create the generalized linear model (GLM) for a binary outcome and the Cox proportional hazards model for time-to-event analysis. The achievement of a 120-h period without reintubation was defined as a successful extubation attempt (EA) within the binary outcome. RESULTS: We evaluated 149 EAs in 81 infants with a median (interquartile range) gestational age of 25+2 (24+3-26+1) weeks. Of this, 90 EAs (60%) were successful while 59 (40%) failed. GLM identified dynamic compliance per kilogram, percentage of spontaneous minute ventilation, and postmenstrual age as significant independent positive variables. Conversely, dynamic compliance variability emerged as a significant independent negative variable for extubation success. This model enabled the creation of a probability estimator for extubation success with a good proportion of sensitivity and specificity (80% and 73% for a cut-off of 60%, respectively). CONCLUSIONS: Ventilator variables reflecting lung mechanical properties and the ability to spontaneously breathe during MV contribute to better prediction of extubation readiness in extremely premature infants with chronic lung disease.

• Klíčová slova
• airway extubation, artificial, chronic lung disease, extremely premature, infant, newborn, respiration,
• MeSH
• extubace * MeSH
• gestační stáří MeSH
• lidé MeSH
• mechanické ventilátory MeSH
• novorozenci extrémně nezralí * MeSH
• novorozenec MeSH
• odpojení od ventilátoru * metody   MeSH
• retrospektivní studie MeSH
• umělé dýchání * metody   MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• pozorovací studie MeSH

The MRE11, RAD50, and NBN genes encode the MRN complex sensing DNA breaks and directing their repair. While carriers of biallelic germline pathogenic variants (gPV) develop rare chromosomal instability syndromes, the cancer risk in heterozygotes remains controversial. We performed a systematic review and meta-analysis of 53 studies in patients with different cancer diagnoses to better understand the cancer risk. We found an increased risk (odds ratio, 95% confidence interval) for gPV carriers in NBN for melanoma (7.14; 3.30-15.43), pancreatic cancer (4.03; 2.14-7.58), hematological tumors (3.42; 1.14-10.22), and prostate cancer (2.44, 1.84-3.24), but a low risk for breast cancer (1.29; 1.00-1.66) and an insignificant risk for ovarian cancer (1.53; 0.76-3.09). We found no increased breast cancer risk in carriers of gPV in RAD50 (0.93; 0.74-1.16; except of c.687del carriers) and MRE11 (0.87; 0.66-1.13). The secondary burden analysis compared the frequencies of gPV in MRN genes in patients from 150 studies with those in the gnomAD database. In NBN gPV carriers, this analysis additionally showed a high risk for brain tumors (5.06; 2.39-9.52), a low risk for colorectal (1.64; 1.26-2.10) and hepatobiliary (2.16; 1.02-4.06) cancers, and no risk for endometrial, and gastric cancer. The secondary burden analysis showed also a moderate risk for ovarian cancer (3.00; 1.27-6.08) in MRE11 gPV carriers, and no risk for ovarian and hepatobiliary cancers in RAD50 gPV carriers. These findings provide a robust clinical evidence of cancer risks to guide personalized clinical management in heterozygous carriers of gPV in the MRE11, RAD50, and NBN genes.

• Klíčová slova
• MRE11, NBN, RAD50, germline variants, meta‐analysis,
• MeSH
• DNA vazebné proteiny genetika   MeSH
• enzymy opravy DNA genetika   MeSH
• genetická predispozice k nemoci * MeSH
• homologní protein MRE11 * genetika   MeSH
• hydrolasy působící na anhydridy kyselin * genetika   MeSH
• jaderné proteiny * genetika   MeSH
• lidé MeSH
• nádory * genetika   MeSH
• proteiny buněčného cyklu * genetika   MeSH
• zárodečné mutace * MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• metaanalýza MeSH
• systematický přehled MeSH
• Názvy látek
• DNA vazebné proteiny MeSH
• enzymy opravy DNA MeSH
• homologní protein MRE11 * MeSH
• hydrolasy působící na anhydridy kyselin * MeSH
• jaderné proteiny * MeSH
• MRE11 protein, human MeSH Prohlížeč
• NBN protein, human MeSH Prohlížeč
• proteiny buněčného cyklu * MeSH
• RAD50 protein, human MeSH Prohlížeč

Gliomagenesis induces profound changes in the composition of the extracellular matrix (ECM) of the brain. In this study, we identified a cellular population responsible for the increased deposition of collagen I and fibronectin in glioblastoma. Elevated levels of the fibrillar proteins collagen I and fibronectin were associated with the expression of fibroblast activation protein (FAP), which is predominantly found in pericyte-like cells in glioblastoma. FAP+ pericyte-like cells were present in regions rich in collagen I and fibronectin in biopsy material and produced substantially more collagen I and fibronectin in vitro compared to other cell types found in the GBM microenvironment. Using mass spectrometry, we demonstrated that 3D matrices produced by FAP+ pericyte-like cells are rich in collagen I and fibronectin and contain several basement membrane proteins. This expression pattern differed markedly from glioma cells. Finally, we have shown that ECM produced by FAP+ pericyte-like cells enhances the migration of glioma cells including glioma stem-like cells, promotes their adhesion, and activates focal adhesion kinase (FAK) signaling. Taken together, our findings establish FAP+ pericyte-like cells as crucial producers of a complex ECM rich in collagen I and fibronectin, facilitating the dissemination of glioma cells through FAK activation.

• Klíčová slova
• collagen type I, extracellular matrix proteins, fibronectin, glioblastoma, pericytes, proteomics,
• MeSH
• endopeptidasy MeSH
• extracelulární matrix * metabolismus   patologie   MeSH
• fibronektiny * metabolismus   MeSH
• glioblastom patologie   metabolismus   MeSH
• gliom * patologie   metabolismus   MeSH
• kolagen typu I metabolismus   MeSH
• lidé MeSH
• membránové proteiny metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí fyziologie   MeSH
• nádory mozku * patologie   metabolismus   MeSH
• pericyty * metabolismus   patologie   MeSH
• pohyb buněk fyziologie   MeSH
• serinové endopeptidasy metabolismus   MeSH
• želatinasy metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• endopeptidasy MeSH
• fibroblast activation protein alpha MeSH Prohlížeč
• fibronektiny * MeSH
• kolagen typu I MeSH
• membránové proteiny MeSH
• serinové endopeptidasy MeSH
• želatinasy MeSH

Mammalian dentition exhibits distinct heterodonty, with more simple teeth located in the anterior area of the jaw and more complex teeth situated posteriorly. While some region-specific differences in signalling have been described previously, here we performed a comprehensive analysis of gene expression at the early stages of odontogenesis to obtain complete knowledge of the signalling pathways involved in early jaw patterning. Gene expression was analysed separately on anterior and posterior areas of the lower jaw at two early stages (E11.5 and E12.5) of odontogenesis. Gene expression profiling revealed distinct region-specific expression patterns in mouse mandibles, including several known BMP and FGF signalling members and we also identified several new molecules exhibiting significant differences in expression along the anterior-posterior axis, which potentially can play the role during incisor and molar specification. Next, we followed one of the anterior molecules, SATB2, which was expressed not only in the anterior mesenchyme where incisor germs are initiated, however, we uncovered a distinct SATB2-positive region in the mesenchyme closely surrounding molars. Satb2-deficient animals demonstrated defective incisor development confirming a crucial role of SATB2 in formation of anterior teeth. On the other hand, ectopic tooth germs were observed in the molar area indicating differential effect of Satb2-deficiency in individual jaw regions. In conclusion, our data provide a rich source of fundamental information, which can be used to determine molecular regulation driving early embryonic jaw patterning and serve for a deeper understanding of molecular signalling directed towards incisor and molar development.

• Klíčová slova
• Incisor, Lower jaw, Microarray, Molar, Mouse, Satb2,
• MeSH
• mandibula * metabolismus   embryologie   MeSH
• myši MeSH
• odontogeneze * genetika   MeSH
• řezáky metabolismus   embryologie   růst a vývoj   MeSH
• rozvržení tělního plánu genetika   MeSH
• signální transdukce MeSH
• stanovení celkové genové exprese * MeSH
• transkripční faktory * genetika   metabolismus   MeSH
• vazebné proteiny DNA v oblastech připojení k matrix * genetika   metabolismus   MeSH
• vývojová regulace genové exprese * MeSH
• zuby metabolismus   embryologie   růst a vývoj   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• SATB2 protein, mouse MeSH Prohlížeč
• transkripční faktory * MeSH
• vazebné proteiny DNA v oblastech připojení k matrix * MeSH

BACKGROUND: Monoallelic germline pathogenic variants (GPVs) in five Fanconi anemia (FA) genes (BRCA1/FANCS, BRCA2/FANCD1, PALB2/FANCN, BRIP1/FANCJ, and RAD51C/FANCO) confer an increased risk of breast (BC) and/or ovarian (OC) cancer, but the role of GPVs in 17 other FA genes remains unclear. METHODS: Here, we investigated the association of germline variants in FANCG/XRCC9 with BC and OC risk. RESULTS: The frequency of truncating GPVs in FANCG did not differ between BC (20/10,204; 0.20%) and OC (8/2966; 0.27%) patients compared to controls (6/3250; 0.18%). In addition, only one out of five tumor samples showed loss-of-heterozygosity of the wild-type FANCG allele. Finally, none of the nine functionally tested rare recurrent missense FANCG variants impaired DNA repair activities (FANCD2 monoubiquitination and FANCD2 foci formation) upon DNA damage, in contrast to all tested FANCG truncations. CONCLUSION: Our study suggests that heterozygous germline FANCG variants are unlikely to contribute to the development of BC or OC.

• Klíčová slova
• Fanconi anemia complementation group G, breast cancer, functional analysis, germline genetic testing, hereditary tumors, ovarian cancer,
• MeSH
• dospělí MeSH
• genetická predispozice k nemoci * MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prsu * genetika   MeSH
• nádory vaječníků * genetika   MeSH
• oprava DNA genetika   MeSH
• protein FANCG * genetika   MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• zárodečné mutace * MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• FANCG protein, human MeSH Prohlížeč
• protein FANCG * MeSH

A mathematical model of myocardial perfusion based on the lattice Boltzmann method (LBM) is proposed and its applicability is investigated in both healthy and diseased cases. The myocardium is conceptualized as a porous material in which the transport and mass transfer of a contrast agent in blood flow is studied. The results of myocardial perfusion obtained using LBM in 1D and 2D are confronted with previously reported results in the literature and the results obtained using the mixed-hybrid finite element method. Since LBM is not suitable for simulating flow in heterogeneous porous media, a simplified and computationally efficient 1D-analog approach to 2D diseased case is proposed and its applicability discussed.

• Klíčová slova
• advection–diffusion problem, contrast agent transport, lattice Boltzmann method, magnetic resonance imaging, mixed‐hybrid finite element method, myocardial perfusion,
• MeSH
• analýza metodou konečných prvků * MeSH
• kontrastní látky MeSH
• koronární cirkulace fyziologie   MeSH
• lidé MeSH
• modely kardiovaskulární * MeSH
• počítačová simulace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kontrastní látky MeSH

Theranostics is a novel paradigm integrating therapy and diagnostics, thereby providing new prospects for overcoming the limitations of traditional treatments. In this context, perfluorocarbons (PFCs) are the most widely used tracers in preclinical fluorine-19 magnetic resonance (19F MR), primarily for their high fluorine content. However, PFCs are extremely hydrophobic, and their solutions often display reduced biocompatibility, relative instability, and subpar 19F MR relaxation times. This study aims to explore the potential of micellar 19F MR imaging (MRI) tracers, synthesized by polymerization-induced self-assembly (PISA), as alternative theranostic agents for simultaneous imaging and release of the non-steroidal antileprotic drug clofazimine. In vitro, under physiological conditions, these micelles demonstrate sustained drug release. In vivo, throughout the drug release process, they provide a highly specific and sensitive 19F MRI signal. Even after extended exposure, these fluoropolymer tracers show biocompatibility, as confirmed by the histological analysis. Moreover, the characteristics of these polymers can be broadly adjusted by design to meet the wide range of criteria for preclinical and clinical settings. Therefore, micellar 19F MRI tracers display physicochemical properties suitable for in vivo imaging, such as relaxation times and non-toxicity, and high performance as drug carriers, highlighting their potential as both diagnostic and therapeutic tools.

• Klíčová slova
• clofazimine, fluorine‐19, magnetic resonance, nanoparticles, polymerization‐induced self‐assembly, theranostics,
• MeSH
• biokompatibilní materiály chemie   MeSH
• fluor chemie   MeSH
• fluorokarbony chemie   MeSH
• halogenace MeSH
• lidé MeSH
• magnetická rezonanční tomografie metody   MeSH
• micely MeSH
• myši MeSH
• nanočástice * chemie   terapeutické užití   MeSH
• teranostická nanomedicína * MeSH
• zobrazování fluorovou magnetickou rezonancí * metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biokompatibilní materiály MeSH
• fluor MeSH
• fluorokarbony MeSH
• micely MeSH

Quantitative phase imaging (QPI) is a powerful tool for label-free visualisation of living cells. Here, we compare two QPI microscopes - the Telight Q-Phase microscope and the Nanolive 3D Cell Explorer-fluo microscope. Both systems provide unbiased information about cell morphology, such as individual cell dry mass, perimeter and area. The Q-Phase microscope uses artefact-free, coherence-controlled holographic imaging technology to visualise cells in real time with minimal phototoxicity. The 3D Cell Explorer-fluo employs laser-based holotomography to reconstruct 3D images of living cells, visualising their internal structures and dynamics. Here, we analysed the strengths and limitations of both microscopes when examining two morphologically distinct cell lines - the cuboidal epithelial MDCK cells which form multicellular clusters and solitary growing Rat2 fibroblasts. We focus mainly on the ability of the devices to generate images suitable for single-cell segmentation by the built-in software, and we discuss the segmentation results and quantitative data generated from the segmented images. We show that both microscopes offer slightly different advantages, and the choice between them depends on the specific requirements and goals of the user.

• Klíčová slova
• MDCK, Rat2, coherence-controlled holographic imaging/microscopy, dry mass content, holotomography, quantitative phase imaging/microscopy, single-cell segmentation,
• MeSH
• buněčné linie MeSH
• holografie * metody   MeSH
• kvantitativní fázové zobrazování MeSH
• lasery MeSH
• mikroskopie * metody   MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: The COVID-19 vaccine was designed to provide protection against infection by the severe respiratory coronavirus 2 (SARS-CoV-2) and coronavirus disease 2019 (COVID-19). However, the vaccine's efficacy can be compromised in patients with immunodeficiencies or the vaccine-induced immunoprotection suppressed by other comorbidity treatments, such as chemotherapy or immunotherapy. To enhance the protective role of the COVID-19 vaccine, we have investigated a combination of the COVID-19 vaccination with ex vivo enrichment and large-scale expansion of SARS-CoV-2 spike glycoprotein-reactive CD4+ and CD8+ T cells. METHODS: SARS-CoV-2-unexposed donors were vaccinated with two doses of the BNT162b2 SARS-CoV-2 vaccine. The peripheral blood mononuclear cells of the vaccinated donors were cell culture-enriched with T cells reactive to peptides derived from SARS-CoV-2 spike glycoprotein. The enriched cell cultures were large-scale expanded using the rapid expansion protocol (REP) and the peptide-reactive T cells were evaluated. RESULTS: We show that vaccination with the SARS-CoV-2 spike glycoprotein-based mRNA COVID-19 vaccine-induced humoral response against SARS-CoV-2 spike glycoprotein in all tested healthy SARS-CoV-2-unexposed donors. This humoral response was found to correlate with the ability of the donors' PBMCs to become enriched with SARS-CoV-2 spike glycoprotein-reactive CD4+ and CD8+ T cells. Using an 11-day REP, the enriched cell cultures were expanded nearly 1000-fold, and the proportions of the SARS-CoV-2 spike glycoprotein-reactive T cells increased. CONCLUSION: These findings show for the first time that the combination of the COVID-19 vaccination and ex vivo T cell large-scale expansion of SARS-CoV-2-reactive T cells could be a powerful tool for developing T cell-based adoptive cellular immunotherapy of COVID-19.

• Klíčová slova
• COVID-19 vaccination, SARS-CoV-2, cellular immunity, ex vivo expansion, humoral immunity, spike glycoprotein-reactive,
• MeSH
• CD8-pozitivní T-lymfocyty imunologie   MeSH
• COVID-19 * imunologie   MeSH
• glykoprotein S, koronavirus imunologie   MeSH
• glykoproteiny MeSH
• leukocyty mononukleární MeSH
• lidé MeSH
• protilátky virové MeSH
• SARS-CoV-2 MeSH
• vakcína BNT162 MeSH
• vakcíny proti COVID-19 * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glykoprotein S, koronavirus MeSH
• glykoproteiny MeSH
• protilátky virové MeSH
• spike protein, SARS-CoV-2 MeSH Prohlížeč
• vakcína BNT162 MeSH
• vakcíny proti COVID-19 * MeSH

Cullin 4B (CUL4B), lysosomal-associated membrane protein Type 2 (LAMP2), ATP1B4, TMEM255A, and ZBTB33 are neighboring genes on Xq24. Mutations in CUL4B result in Cabezas syndrome (CS). Male CS patients present with dysmorphic, neuropsychiatric, genitourinary, and endocrine abnormalities. Heterozygous CS females are clinically asymptomatic. LAMP2 mutations cause Danon disease (DD). Cardiomyopathy is a dominant feature of DD present in both males and heterozygous females. No monogenic phenotypes have been associated with mutations in ATP1B4, TMEM255A, and ZBTB33 genes. To facilitate diagnostics and counseling in CS and DD families, we present a female DD patient with a de novo Alu-mediated Xq24 rearrangement causing a deletion encompassing CUL4B, LAMP2, and also the other three neighboring genes. Typical to females heterozygous for CUL4B mutations, the patient was CS asymptomatic, however, presented with extremely skewed X-chromosome inactivation (XCI) ratios in peripheral white blood cells. As a result of the likely selection against CUL4B deficient clones, only minimal populations (~3%) of LAMP2 deficient leukocytes were identified by flow cytometry. On the contrary, myocardial LAMP2 protein expression suggested random XCI. We demonstrate that contiguous CUL4B and LAMP2 loss-of-function copy number variations occur and speculate that male patients carrying similar defects could present with features of both CS and DD.

• Klíčová slova
• Cabezas syndrome, Danon disease, cullin 4B, female heterozygotes, lysosomal-associated membrane protein 2,
• MeSH
• chromozomální delece MeSH
• dospělí MeSH
• elementy Alu genetika   MeSH
• exony genetika   MeSH
• glykogenóza typu IIb diagnóza   genetika   patofyziologie   MeSH
• inaktivace chromozomu X genetika   MeSH
• kardiomyopatie genetika   patofyziologie   MeSH
• kulinové proteiny genetika   MeSH
• lidé MeSH
• membránový protein 2 asociovaný s lyzozomy genetika   MeSH
• mentální retardace vázaná na chromozom X genetika   patofyziologie   MeSH
• mutace ztráty funkce genetika   MeSH
• myokard metabolismus   MeSH
• sodíko-draslíková ATPasa genetika   MeSH
• transkripční faktory genetika   MeSH
• variabilita počtu kopií segmentů DNA genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• práce podpořená grantem MeSH
• Názvy látek
• ATP1B4 protein, human MeSH Prohlížeč
• CUL4B protein, human MeSH Prohlížeč
• kulinové proteiny MeSH
• LAMP2 protein, human MeSH Prohlížeč
• membránový protein 2 asociovaný s lyzozomy MeSH
• sodíko-draslíková ATPasa MeSH
• transkripční faktory MeSH