BACKGROUND: The canonical Wnt signaling pathway controls the continuous renewal of the intestinal epithelium and the specification of epithelial cell lineages. Tcf4, a nuclear mediator of Wnt signaling, is essential for the differentiation and maintenance of Paneth cells in the small intestine. Its deficiency is associated with reduced expression of key α-defensins, highlighting its role in host-microbe interactions. However, the exact function of Tcf4 in specifying the secretory lineage and its contribution to antimicrobial peptide production remain incompletely understood. Remarkably, α-defensin expression has also been detected in human colon adenomas, where aberrant Wnt signaling is a hallmark. This raises important questions: What is the role of these Paneth-like cells in tumor biology, and how does Tcf4 influence their identity and function? METHODS: We investigated cell specification in small intestinal crypts and colon tumors using conditional Tcf7l2 deletion, cell type-specific Cre recombinases, and reporter alleles in mice. Transcriptomic (single-cell and bulk RNA sequencing) and histological analyses were performed and complemented by microbiome profiling, antibiotic treatment, and intestinal organoids to functionally validate the main findings. RESULTS: The inactivation of Tcf4 depletes Paneth cells and antimicrobial peptides, disrupting the gut microbiota balance. In secretory progenitors, loss of Tcf4 shifts differentiation toward goblet cells. In the small intestine, alternative secretory progenitors produce Wnt ligands to support stem cells and epithelial renewal in the absence of Paneth cells. In colon tumors, Paneth-like cells form a tumor cell population, express Wnt ligands, and require Tcf4 for their identity. Loss of Tcf4 redirects their differentiation toward goblet cells. CONCLUSIONS: Tcf4 controls the balance between Paneth and goblet cells and is essential for antimicrobial peptide production in the small intestine. In colon adenomas, Paneth-like tumor cells drive antimicrobial gene expression and provide Wnt3 ligands, which may have implications for cancer therapy.
- MeSH
- alfa-defensiny metabolismus MeSH
- buněčná diferenciace MeSH
- lidé MeSH
- myši MeSH
- nádory tračníku * patologie genetika mikrobiologie metabolismus MeSH
- organoidy metabolismus MeSH
- Panethovy buňky metabolismus MeSH
- pohárkové buňky metabolismus MeSH
- signální dráha Wnt MeSH
- střevní mikroflóra * MeSH
- tenké střevo * metabolismus patologie mikrobiologie MeSH
- transkripční faktor 4 * metabolismus genetika MeSH
- transkriptom * MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
This review aims at the current trends in chocolate ganache production and recipe formulation. Ganache is a blend of chocolate, sugars, dairy, and other ingredients commonly used to fill pralines, pastries, etc. In spite of ganache's popularity in the food industry, a comprehensive review focused on the application of functional substances and ganache processing has not been discussed in the scientific literature. This review addresses the new ways of applying special ingredients, such as vegetable fats and seeds, flavor infusions, oleogels, hemp products, etc., which can be added to the ganache matrix to achieve desirable properties. In particular, the application of sterols and sterol esters as functional substances of oleogels seems to be a very promising method, enhancing the ganache fat profile. The elevated caloric content that is characteristic of ganache can be substantially attenuated through the application of hydrocolloids and/or fruit-based components, thereby offering the potential for caloric reduction without compromising on taste. The various alterations to ganache formulations by the application of natural substances offer a large base for the development of novel ganache variants and relevant food products.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
In the past decade, single-cell transcriptomics has helped to uncover new cell types and states and led to the construction of a cellular compendium of health and disease. Despite this progress, some difficult-to-sequence cells remain absent from tissue atlases. Eosinophils-elusive granulocytes that are implicated in a plethora of human pathologies1-5-are among these uncharted cell types. The heterogeneity of eosinophils and the gene programs that underpin their pleiotropic functions remain poorly understood. Here we provide a comprehensive single-cell transcriptomic profiling of mouse eosinophils. We identify an active and a basal population of intestinal eosinophils, which differ in their transcriptome, surface proteome and spatial localization. By means of a genome-wide CRISPR inhibition screen and functional assays, we reveal a mechanism by which interleukin-33 (IL-33) and interferon-γ (IFNγ) induce the accumulation of active eosinophils in the inflamed colon. Active eosinophils are endowed with bactericidal and T cell regulatory activity, and express the co-stimulatory molecules CD80 and PD-L1. Notably, active eosinophils are enriched in the lamina propria of a small cohort of patients with inflammatory bowel disease, and are closely associated with CD4+ T cells. Our findings provide insights into the biology of eosinophils and highlight the crucial contribution of this cell type to intestinal homeostasis, immune regulation and host defence. Furthermore, we lay a framework for the characterization of eosinophils in human gastrointestinal diseases.
- MeSH
- analýza genové exprese jednotlivých buněk MeSH
- antigeny CD80 metabolismus MeSH
- eozinofily * klasifikace cytologie imunologie metabolismus MeSH
- idiopatické střevní záněty imunologie MeSH
- imunita * MeSH
- interferon gama MeSH
- interleukin 33 MeSH
- kolitida * imunologie patologie MeSH
- lidé MeSH
- myši MeSH
- proteom MeSH
- střeva * imunologie patologie MeSH
- T-lymfocyty MeSH
- transkriptom MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Helicobacter pylori infection is a major risk factor for the development of gastric cancer. The bacteria reside in close proximity to gastric surface mucous as well as stem and progenitor cells. Here, we take advantage of wild-type and genetically engineered murine gastric organoids and organoid-derived monolayers to study the cellular targets of H. pylori-induced DNA damage and replication stress and to explore possible interactions with preexisting gastric cancer driver mutations. We find using alkaline comet assay, single-molecule DNA fiber assays, and immunofluorescence microscopy of DNA repair foci that H. pylori induces transcription-dependent DNA damage in actively replicating, Leucine-rich-repeat containing G-Protein-Coupled Receptor 5 (Lgr5)-positive antral stem and progenitor cells and their Troy-positive corpus counterparts, but not in other gastric epithelial lineages. Infection-dependent DNA damage is aggravated by Apc inactivation, but not by Trp53 or Smad4 loss, or Erbb2 overexpression. Our data suggest that H. pylori induces DNA damage in stem and progenitor cells, especially in settings of hyperproliferation due to constitutively active Wnt signaling.
- MeSH
- Helicobacter pylori * genetika MeSH
- infekce vyvolané Helicobacter pylori * genetika mikrobiologie MeSH
- kmenové buňky MeSH
- lidé MeSH
- myši MeSH
- nádory žaludku * patologie MeSH
- poškození DNA MeSH
- receptory spřažené s G-proteiny genetika MeSH
- tumor supresorové geny MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
Úvod: Krční emfyzém bývá jedním z průvodních příznaků poranění polykacích/ dýchacích cest nebo přechází na krk z jiných oblastí. V prvních hodinách po jeho vzniku může být klinický obraz dramatický, provázený dušností, odyno-/ dysfagií, případně bolestí na hrudi nebo mezi lopatkama. V diferenciální diagnostice je kromě traumatické etiologie třeba vyloučit i další akutní stavy, např. akutní koronární syndrom aj. Emfyzém bývá různého rozsahu, často je provázen pneumomediastinem a/nebo plášťovým pneumotoraxem. Publikace si klade za cíl především diagnostickou a terapeutickou rozvahu, nikoliv zjištění incidence emfyzému. Metodika: Jedná se o retrospektivní studii z ORL pracovišť Pardubické nemocnice, Fakultní nemocnice Hradec Králové a Nemocnice Liberec za období 15 let (2005–2020). Sledovány byly pouze klinicky významné emfyzémy, emfyzémy nejasné etiologie a emfyzémy s nutností intenzivní péče. Výsledky: V souboru 15 nemocných jsou následující příčiny krčního emfyzému – 3 úrazy hrtanu, 3 Hammanovy syndromy, 2 tracheální intubace, 2 úrazy polykacích cest, 2 onemocnění covid-19, 1 tonzilektomie, 1 urgentní tracheostomie a 1 kýchnutí. Etiologicky je významným (spolu)faktorem Valsalvův manévr, který dominuje zejména u mladistvých a mladých dospělých (53 %, 8 pacientů). Úrazová etiologie se vyskytovala ve věku 17–53 let (33 %, 5 pacientů). Iatrogenní postižení je častější u pacientů nad 55 let. Pneumomediastinum se vyskytlo u 47 %, plášťový pneumotorax u 40 % nemocných. Čtyři zpočátku diagnosticky nejasné případy rozebíráme podrobněji. Závěr: U nemocného s krčním emfyzémem jsou indikována zobrazovací a endoskopická vyšetření. V terapii je vždy nutné profylaktické nasazení širokospektrých antibiotik, observace na lůžku a mezioborová spolupráce.
Introduction: Emphysema of the neck represents a crucial symptom of swallowing/respiratory pathways injuries or rarely passes to the neck area from different locations. The clinical image can develop dramatically in the first hours after onset and frequently the etiology remains unclear. Dyspnoea, dysphagia, retrosternal or interscapular pain usually follows. In the differential diagnosis, not only traumatic etiology but also other acute conditions must be considered – e. g. acute coronary syndrome etc. The size of emphysema varies, also head and chest area can also be affected. Pneumomediastinum or partial pneumothorax are also often presented. The primarily aim of this paper is to discuss a diagnostic and therapeutic possibilities, not to determine the incidence of emphysema. Methodology: This is a retrospective multicenter study involving patients from three regional ENT departments – Pardubice Hospital, Hradec Králové University Hospital and Liberec Hospital over a period of 15 years (2005–2020). Only clinically significant emphysema, emphysema of unclear origin and emphysema requiring intensive care were in-rolled. Results: In a group of 15 patients, there are the following causes of cervical emphysema – 3 laryngeal injuries, 3 Hamman’s syndromes, 2 tracheal intubations, 2 traumas of swallowing pathways, 2 in COVID-19 disease, 1 tonsillectomy, 1 urgent tracheostomy and 1 sneezing. Etiologically, the Valsalva maneuver is an important (co)factor, dominating especially in adolescents and young adults (53%, 8 patients). Traumatic etiology occurred at the age of 17–53 years (33%, 5 patients). Iatrogenic involvement is more common over the age of 55 years. Pneumomediastinum occurred in 47% and partial pneumothorax in 40% of patients. Four of those initially unclear cases are discussed in detail. Conclusion: Imaging methods and endoscopic examination are indicated in a patient with cervical emphysema of unclear origin. Prophylactic use of broad-spectrum antibio tics, observation and interdisciplinary cooperation are always necessary.
- MeSH
- COVID-19 komplikace MeSH
- emfyzém * diagnóza etiologie terapie MeSH
- Hammanův-Richův syndrom komplikace MeSH
- lidé MeSH
- mediastinální emfyzém komplikace MeSH
- pneumotorax komplikace MeSH
- poškození plic komplikace MeSH
- retrospektivní studie MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
BACKGROUND: This study focuses on the hearing threshold for bone conduction (BC) after middle-ear surgery. METHODS: A total of 92 patients (120 ears) were treated for newly diagnosed chronic otitis media with cholesteatoma (2013-2018). BC was examined at frequencies of 0.5, 1, 2, and 4 kHz prior to and 1 year after surgery. STAM classification for cholesteatoma location, EAONO/JOS for stage, and surgery according to SAMEO-ATO classification were applied. The bone conduction threshold was compared for individual frequencies in patients with occurrence/absence of cholesteatoma in different locations. RESULTS: For the occurrence of cholesteatoma in the attic (A), a statistically significant difference was found at 4 kHz (p < 0.001), in the supratubal recess (S1) at 4 kHz (p = 0.003), and for the mastoid (M) at 0.5 kHz (p = 0.024), at 1 kHz (p = 0.032), and at 2 kHz (p = 0.039). CONCLUSIONS: Cholesteatoma location can influence the post-operative hearing threshold for bone conduction.
- Publikační typ
- časopisecké články MeSH
Poranění hrtanu je málo častý stav, který může ohrozit pacienta na životě obstrukcí dýchacích cest. Časná diagnostika je klíčovým faktorem v jeho řešení. K hodnocení rozsahu postižení se nejčastěji používá Schaeferova klasifikace, podle které vznikl doporučený postup terapie. Názory na zajištění dýchacích cest orotracheální intubací v přednemocniční péči se v literatuře různí. Autoři prezentují případ 40letého muže se zlomeninou štítné chrupavky vzniklé po úderu pěstí do krku. Klinicky byl pacient bez dušnosti, byl přítomen otok měkkých tkání krku a podkožní emfyzém. Zobrazovací vyšetření prokázalo frakturu kostry hrtanu. V celkové anestezii byla provedena revize a rekonstrukce štítné chrupavky a zajišťovací tracheostomie. Pooperační průběh byl příznivý a pacient byl dekanylován. Při zevních poraněních krku je důležité pomýšlet na poškození struktur hrtanu. Časná diagnostika a terapie jsou klíčové, zásadní význam mají v první řadě zabezpečení dýchacích cest a prevence dušení, dále je snaha zajistit předpoklady pro zachování funkcí hrtanu (dýchání, fonace a polykání).
Laryngeal injury is a rare condition that may threaten the patient's life with airway obstruction. Early diagnostics is the key management factor. Schaefer's classification is the most commonly used classification to assess the extent of the injury. Based on this classification, a recommended management of therapy has been developed. There are several opinions on securing the airway by the orotracheal intubation. The authors present a case report of a 40-year-old male with thyroid cartilage fracture caused by a punch to the neck. The patient did not show signs of dyspnoea, had swollen neck and subcutaneous emphysema. Imaging by computed tomography showed a laryngeal fracture. A reconstruction of the thyroid cartilage and a tracheostomy in general anaesthesia were performed. Postoperative course was propitious, and the patient could be decannulated. It is important to consider a laryngeal injury in patients with external neck trauma. The key elements are early diagnosis and therapy, in which securing the airway is the crucial step. Further an effort should be made to create conditions for preserving laryngeal functions (respiration, phonation, and swallowing).
We lack a holistic understanding of the genetic programs orchestrating embryonic colon morphogenesis and governing damage response in the adult. A window into these programs is the transcriptomes of the epithelial and mesenchymal cell populations in the colon. Performing unbiased single-cell transcriptomic analyses of the developing mouse colon at different embryonic stages (embryonic day 14.5 [E14.5], E15.5, and E18.5), we capture cellular and molecular profiles of the stages before, during, and after the appearance of crypt structures, as well as in a model of adult colitis. The data suggest most adult lineages are established by E18.5. We find embryonic-specific gene expression profiles and cell populations that reappear in response to tissue damage. Comparison of the datasets from mice and human colitis suggests the processes are conserved. In this study, we provide a comprehensive single-cell atlas of the developing mouse colon and evidence for the reactivation of embryonic genes in disease.
- MeSH
- analýza jednotlivých buněk MeSH
- buněčná diferenciace MeSH
- embryo savčí metabolismus MeSH
- idiopatické střevní záněty genetika patologie MeSH
- kolitida genetika MeSH
- kolon embryologie patologie MeSH
- lidé MeSH
- mezoderm embryologie MeSH
- modely nemocí na zvířatech MeSH
- myši inbrední C57BL MeSH
- stanovení celkové genové exprese * MeSH
- střevní sliznice embryologie metabolismus patologie MeSH
- vývojová regulace genové exprese MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Úvod: Nově vzniklý systém SAMEO-ATO (podle konsenzu IOOG, 2018) popisuje klasifikaci středoušních operací. Navazuje tak na konsenzus definice, klasifikace a stagingu středoušního cholesteatomu (EAONO a JOS, 2017). Metodika: Podle klasifikace SAMEO-ATO jsme retrospektivně oklasifikovali soubor pacientů operovaných pro chronický středoušní zánět s cholesteatomem v letech 2013–2017. Výsledky: Ve sledovaném období bylo pro chronický hnisavý středoušní zánět s cholesteatomem operováno 104 pacientů – 56 mužů a 48 žen ve věkovém rozmezí 5–77 let s věkovým průměrem 33 a mediánem 27 let. Celkem bylo provedeno 142 operačních výkonů, z nichž bylo 77 primárních (S1), 40 plánovaných kontrolních "second look" operací (S2p) a 25 operací bylo revizních (S2r). U plánovaných "second look" operací (S2p) byl prokázán reziduální cholesteatom u 20 (50 %) pacientů, u neplánovaných revizních operací (S2r) byla prokázána recidiva cholesteatomu u 19 (76 %) pacientů. Práce se dále zabývá zastoupením provedených výkonů v každé z kategorií klasifikace. V případě mastoidektomie (parametr “M”) byl nejčastěji prováděn typ M1a + 2a (N = 49; 34,51 %), dále M2c (N = 25; 17,61 %), M2a (N = 20; 14,08 %), M1a (N = 4; 2,82 %), M2b (N = 5; 3,52 %), M3a (N = 3; 2,11 %) a M3b (N = 1; 0,70 %). V případě rekonstrukcí řetězu kůstek (parametr “O”) byl nejčastějším typem výkonu Osd (N = 58; 40,85 %), dále Ost (N = 21; 14,79 %), Oft (N = 19; 13,38 %), Ofd (N = 5; 3,52 %) a Osm (N = 5; 3,52 %). Závěr: Systém SAMEO-ATO zahrnuje jednotlivé aspekty chirurgických technik včetně rekonstrukce převodního systému a umožňuje vytvoření jednotné databáze pacientů po středoušních operacích. Metoda se může uplatnit v porovnávání chirurgických výsledků při použití různých operačních technik, sledování výskytu cholesteatomu a výsledků operací v rámci pracovišť včetně možného využití dat k výzkumným účelům.
Introduction: The SAMEO-ATO system established by IOOG consensus in 2018 introduces a new, complex, and unified classification of middle ear surgery. This system is generally suitable for various surgical procedures in the middle ear. The main goal of this work is to present our experience of using the new classification system in the clinical practice focusing on cholesteatoma surgery. Methods: We have retrospectively classified the group of patients who underwent surgery for chronic otitis media with cholesteatoma between 2013 and 2017. Results: We classified 142 cholesteatoma surgeries carried out on 104 patients (56 men and 48 women at the age 5–77 years, with the mean 33 years and the median 27 years) using the SAMEO-ATO classification. There were 77 primary surgeries (S1), 40 second-look surgeries (S2p) and 25 secondary surgeries for recidivism (S2r). There was a 50% (20) incidence of recurrent cholesteatoma in S2p surgeries and 76% (19) incidence of residual cholesteatoma in S2r. We have further evaluated our group of patients by each SAMEO-ATO subclassification. In the mastoid part of surgery (parameter “M”), the combination of M1a + 2a (N = 49, 34.51%) was most frequent, followed by M2c (N = 25, 17.61%), M2a (N = 20, 14.08%), M1a (N = 4, 2.82%), M2b (N = 5, 3.52%), M3a (N = 3, 2.11%) and M3b (N = 1, 0.70%). In the ossicular reconstruction (parameter “O”), the most frequent was Osd (N = 58, 40.85%), followed by Ost (N = 21, 14.79%), Oft (N = 19, 13.38%), Ofd (N = 5, 3.52%) and Osm (N = 5, 3.52%). Conclusions: The SAMEO-ATO classifies various surgical techniques of cholesteatoma removal as well as reconstruction of the ossicular chain. It allows the creation of a unified database of patients who underwent middle ear surgery and the comparison of the results of different surgery techniques among various departments. The collected data are applicable for research purposes.
- Klíčová slova
- SAMEO-ATO,
- MeSH
- cholesteatom středního ucha * chirurgie klasifikace MeSH
- lidé MeSH
- otologické chirurgické výkony * klasifikace MeSH
- retrospektivní studie MeSH
- Check Tag
- lidé MeSH
The homeostasis of the gut epithelium relies upon continuous renewal and proliferation of crypt-resident intestinal epithelial stem cells (IESCs). Wnt/β-catenin signaling is required for IESC maintenance, however, it remains unclear how this pathway selectively governs the identity and proliferative decisions of IESCs. Here, we took advantage of knock-in mice harboring transgenic β-catenin alleles with mutations that specifically impair the recruitment of N- or C-terminal transcriptional co-factors. We show that C-terminally-recruited transcriptional co-factors of β-catenin act as all-or-nothing regulators of Wnt-target gene expression. Blocking their interactions with β-catenin rapidly induces loss of IESCs and intestinal homeostasis. Conversely, N-terminally recruited co-factors fine-tune β-catenin's transcriptional output to ensure proper self-renewal and proliferative behaviour of IESCs. Impairment of N-terminal interactions triggers transient hyperproliferation of IESCs, eventually resulting in exhaustion of the self-renewing stem cell pool. IESC mis-differentiation, accompanied by unfolded protein response stress and immune infiltration, results in a process resembling aberrant "villisation" of intestinal crypts. Our data suggest that IESC-specific Wnt/β-catenin output requires selective modulation of gene expression by transcriptional co-factors.
- MeSH
- algoritmy MeSH
- beta-katenin chemie metabolismus MeSH
- buněčná diferenciace MeSH
- chromatin metabolismus MeSH
- fenotyp MeSH
- genetická transkripce * MeSH
- homeostáza MeSH
- hyperplazie MeSH
- JNK mitogenem aktivované proteinkinasy metabolismus MeSH
- kmenové buňky metabolismus MeSH
- messenger RNA genetika metabolismus MeSH
- mutace genetika MeSH
- mutantní proteiny metabolismus MeSH
- myši MeSH
- organoidy metabolismus MeSH
- proliferace buněk MeSH
- restrukturace chromatinu MeSH
- sekvence nukleotidů MeSH
- signální transdukce MeSH
- střevní sliznice cytologie MeSH
- transkripční faktory metabolismus MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
