The diagnosis of solid tumors of epithelial origin (carcinomas) represents a major part of the workload in clinical histopathology. Carcinomas consist of malignant epithelial cells arranged in more or less cohesive clusters of variable size and shape, together with stromal cells, extracellular matrix, and blood vessels. Distinguishing stroma from epithelium is a critical component of artificial intelligence (AI) methods developed to detect and analyze carcinomas. In this paper, we propose a novel automated workflow that enables large-scale guidance of AI methods to identify the epithelial component. The workflow is based on re-staining existing hematoxylin and eosin (H&E) formalin-fixed paraffin-embedded sections by immunohistochemistry for cytokeratins, cytoskeletal components specific to epithelial cells. Compared to existing methods, clinically available H&E sections are reused and no additional material, such as consecutive slides, is needed. We developed a simple and reliable method for automatic alignment to generate masks denoting cytokeratin-rich regions, using cell nuclei positions that are visible in both the original and the re-stained slide. The registration method has been compared to state-of-the-art methods for alignment of consecutive slides and shows that, despite being simpler, it provides similar accuracy and is more robust. We also demonstrate how the automatically generated masks can be used to train modern AI image segmentation based on U-Net, resulting in reliable detection of epithelial regions in previously unseen H&E slides. Through training on real-world material available in clinical laboratories, this approach therefore has widespread applications toward achieving AI-assisted tumor assessment directly from scanned H&E sections. In addition, the re-staining method will facilitate additional automated quantitative studies of tumor cell and stromal cell phenotypes.

• MeSH
• barvení a značení MeSH
• deep learning * MeSH
• eosin MeSH
• epitelové buňky MeSH
• hematoxylin MeSH
• keratiny * MeSH
• lidé MeSH
• umělá inteligence MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Genes influencing oocyte maturation may be valuable for predicting their developmental potential, as well as discerning the mechanistic pathways regulating oocyte development. In the presented research microarray gene expression analysis of immature and in vitro matured porcine oocytes was performed. Two groups of oocytes were compared in the study: before (3 × n = 50) and after in vitro maturation (3 × n = 50). The selection of viable oocytes was performed using the brilliant cresyl blue (BCB) test. Furthermore, microarrays and RT-qPCR was used to analyze the transcriptome of the oocytes before and after IVM. The study focused on the genes undergoing differential expression in two gene-ontology groups: "Cellular response to hormone stimulus" and "Cellular response to unfolded protein", which contain genes that may directly or indirectly be involved in signal transduction during oocyte maturation. Examination of all the genes of interest showed a lower level of their expression after IVM. From the total number of genes in these gene ontologies ten of the highest change in expression were identified: FOS, ID2, BTG2, CYR61, ESR1, AR, TACR3, CCND2, EGR2 and TGFBR3. The successful maturation of the oocytes was additionally confirmed with the use of lipid droplet assay. The genes were briefly described and related to the literature sources, to investigate their potential roles in the process of oocyte maturation. The results of the study may serve as a basic molecular reference for further research aimed at improving the methods of oocyte in vitro maturation, which plays an important role in the procedures of assisted reproduction.

• MeSH
• eosin chemie   MeSH
• hematoxylin chemie   MeSH
• hormony genetika   metabolismus   MeSH
• IVM techniky * MeSH
• kultivované buňky MeSH
• lipidy analýza   MeSH
• oocyty růst a vývoj   metabolismus   MeSH
• oxaziny chemie   MeSH
• prasata MeSH
• signální transdukce MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

The mucosa of uterine tube forms multiple and branched longitudinal mucosal folds and takes part in many reproduction events, such as oocyte pick-up, gamete transport, sperm capacitation, fertilization, and early embryonic development. In the habilitation thesis of German physician Paul Kroemer (1904) was the first to describe the lymphatic lacunae inside the tubal folds (by injection of Indian ink), which the author named the öLymphbahnen" (ölymphatic channels"). Despite the fact that this first description has existed for 110 years, there is no mention of these lacunae in most of the current literature. In this article we present a rediscovery of completely overlooked morphological structures of uterine tubes - the lymphatic lacunae in their mucosal folds. The specimens from the uterine tubes were taken from 72 women (mean age 46.25 years) who underwent transabdominal or laparoscopic salpingectomy. The tissue samples from anatomically different parts of the uterine tubes were used for hematoxylin and eosin staining and for immunohistochemistry. Primary antibodies were used to label and detect podoplanin D2-40, a selective marker of lymphatic endothelia, CD34 antigen, and von Willebrand factor (Factor VIII). In the histological slides of the uterine tubes, there were noticeable slits or gaps within the loose connective tissue of the lamina propria of the mucosal folds. They were lined with one layer of squamous endothelial cells. These öempty spaces" were most prominent in the fimbriae, but were still well recognizable in mucosal folds of the ampulla. They always run through the central part of the fold. As a results of immunohistochemistry, we confirmed that in the centre of every mucosal fold, as well as in the fimbriae of the uterine tubes, dilated lymphatic spaces were situated and were lined with a simple layer of lymphatic endothelial cells (positive for podoplanin and CD34, and negative for Factor VIII). As there is no mention on them in the current Terminologia Histologica, we proposed the term ölymphatic lacunae of tubal mucosal folds and fimbriae" in English and ölacunae lymphaticae plicae mucosae et fimbriae" in Latin. According to our hypothesis, these lymphatic lacunae may be responsible for the thickening of the fimbriae during the oocyte pick-up and the maintenance of the tubal fluid.

• MeSH
• barvicí látky MeSH
• dospělí MeSH
• eosin MeSH
• hematoxylin MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfoidní tkáň cytologie   MeSH
• salpingektomie MeSH
• vejcovody anatomie a histologie   fyziologie   chirurgie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Úvod: Česká republika se dlouhodobě řadí ke státům s nejvyšším výskytem kolorektálního karcinomu. Při vývoji kolorektální neoplazie hraje významnou roli proliferace, diferenciace a apoptóza buněk. Cílem naší studie bylo stanovit míru apoptózy u jednotlivých stadií kolorektální neoplazie. Metodika: Apoptóza byla stanovena ve slizničních biopsiích kolorektální neoplazie a normální sliznice u 20 pacientů s nepokročilým kolorektálním adenomem, u 20 pacientů s pokročilým kolorektálním adenomem, u 20 pacientů s kolorektálním karcinomem a u 20 jedinců s normálním nálezem v kolorektu. Míra apoptózy byla hodnocena po obarvení hematoxylin-eozinem (ve třech kompartmentech: povrchový kompartment, horní a dolní část krypt) a imunohistochemickými metodami (detekcí aktivované kaspázy-3). Výsledky: Apoptotická aktivita byla vyjádřena pomocí apoptotického indexu. Ve zdravé sliznici kolorekta byla pozorována v dolní části krypt nízká úroveň apoptotické aktivity. V horní části krypt klesla apoptotická aktivita k nulovým hodnotám. Nárůst apoptotické aktivity byl pozorován v povrchovém kompartmentu. Přeměna zdravé sliznice v tkáň nepokročilého kolorektálního adenomu byla doprovázena statisticky signifikantním vzestupem apoptotické aktivity ve všech třech kompartmentech (p ≤ 0,05), přičemž nejvyšší nárůst byl pozorován v horní části krypt. Přeměna nepokročilého adenomu v pokročilý byla spojena s dalším navýšením apoptotické aktivity, a to opět především v horní části krypt. U karcinomů ve srovnání s pokročilým adenomem došlo k významnému poklesu apoptotické aktivity ve všech třech kompartmentech (p ≤ 0,05). Výsledky imunohistochemických hodnocení potvrdily trend pozorovaný u vzorků barvených hematoxylin-eozinem. Závěry: Prokázali jsme přítomnost deregulace apoptózy při vývoji kolorektální neoplazie. Ovlivnění míry apoptózy především při přeměně pokročilého adenomu v kolorektální karcinom by mělo v klinické praxi příznivé konsekvence.

Background: The Czech Republic has one of the highest incidence rates of colorectal cancer. Cell proliferation, differentiation, and apoptosis play a key role in the development of this disease. The aim of our study was to assess the level of apoptosis at each stage of colorectal neoplasia. Methodology: Apoptosis was evaluated by examining mucosal biopsies of colorectal neoplasm and normal mucosa. This was performed in 20 patients with non-advanced adenoma, 20 patients with advanced adenoma, 20 patients with colorectal carcinoma, and 20 individuals with normal colorectal findings. The grade of apoptosis was assessed after hematoxylin-eosin staining (in three compartments: the superficial compartment and the upper and lower parts of the crypts) and by immunohistochemical methods (by detection of activated-caspase-3). Results: Apoptotic activity was reported as an apoptotic index. In healthy colorectal mucosa, low apoptotic activity was observed in the lower part of the crypts. In the upper part of the crypts, apoptotic activity decreased (to almost zero) and in the superficial compartment, there was an increase in apoptotic activity. Transformation of healthy mucosa into non-advanced colorectal adenoma was associated with a statistically significant increase in apoptotic activity in all three compartments (p ≤ 0.05), with the biggest increase in the upper part of the crypts. Transformation of non-advanced adenoma into advanced adenoma was related to further increases in apoptotic activity, again, mainly in the upper part of the crypts. There was a statistically significant decrease in apoptotic activity in all three comparments of carcinoma samples compared to advanced adenoma (p ≤ 0.05). Results of immunohistochemical methods confirmed this trend. Conclusions: We have shown deregulation of apoptosis during the development of colorectal neoplasia. Being able to influence the degree of apoptosis, especially during the transformation of an advanced adenoma into a carcinoma, would have beneficial consequences in clinical practice.

• Klíčová slova
• kolorektální adenom,
• MeSH
• adenom * metabolismus   patologie   MeSH
• apoptóza * MeSH
• biopsie MeSH
• dospělí MeSH
• eosin MeSH
• hematoxylin MeSH
• imunohistochemie MeSH
• karcinogeneze MeSH
• karcinom metabolismus   patologie   MeSH
• kolorektální nádory * metabolismus   patologie   MeSH
• neparametrická statistika MeSH
• střevní sliznice metabolismus   patologie   MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• Publikační typ
• práce podpořená grantem MeSH

Shock waves can cause significant cytotoxic effects in tumor cells and tissues both in vitro and in vivo. However, understanding the mechanisms of shock wave interaction with tissues is limited. We have studied in vivo effects of focused shock waves induced in the syngeneic sarcoma tumor model using the TUNEL assay, immunohistochemical detection of caspase-3 and hematoxylin-eosin staining. Shock waves were produced by a multichannel pulsed-electrohydraulic discharge generator with a cylindrical ceramic-coated electrode. In tumors treated with shock waves, a large area of damaged tissue was detected which was clearly differentiated from intact tissue. Localization and a cone-shaped region of tissue damage visualized by TUNEL reaction apparently correlated with the conical shape and direction of shock wave propagation determined by high-speed shadowgraphy. A strong TUNEL reaction of nuclei and nucleus fragments in tissue exposed to shock waves suggested apoptosis in this destroyed tumor area. However, specificity of the TUNEL technique to apoptotic cells is ambiguous and other apoptotic markers (caspase-3) that we used in our study did not confirmed this observation. Thus, the generated fragments of nuclei gave rise to a false TUNEL reaction not associated with apoptosis. Mechanical stress from high overpressure shock wave was likely the dominant pathway of tumor damage.

• MeSH
• design vybavení MeSH
• elektrostimulační terapie přístrojové vybavení   metody   MeSH
• eosin MeSH
• experimentální nádory patologie   terapie   MeSH
• fluorescence MeSH
• hematoxylin MeSH
• imunohistochemie metody   MeSH
• kaspasa 3 metabolismus   MeSH
• koncové značení zlomů DNA in situ MeSH
• potkani inbrední LEW MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Combination of fluorescence techniques and molecular docking was used to monitor interaction of Na,K-ATPase and its large cytoplasmic loop connecting fourth and fifth transmembrane helices (C45) with fluorone dyes (i.e. eosin Y, 5(6)-carboxyeosin, rose bengal, fluorescein, and erythrosine B). Our data suggested that there are at least two binding sites for all used fluorone dyes, except of 5(6)-carboxyeosin. The first binding site is located on C45 loop, and it is sensitive to the presence of nucleotide. The other site is located on the extracellular part of the enzyme, and it is sensitive to the presence of Na(+) or K(+) ions. The molecular docking revealed that in the open conformation of C45 loop (which is obtained in the presence of ATP) all used fluorone dyes occupy position directly inside the ATP-binding pocket, while in the closed conformation (i.e. in the absence of any ligand) they are located only near the ATP-binding site depending on their different sizes. On the extracellular part of the protein, the molecular docking predicts two possible binding sites with similar binding energy near Asp897(α) or Gln69(β). The former was identified as a part of interaction site between α- and β-subunits, the latter is in contact with conserved FXYD sequence of the γ-subunit. Our findings provide structural explanation for numerous older studies, which were performed with fluorone dyes before the high-resolution structures were known. Further, fluorone dyes seem to be good probes for monitoring of intersubunit interactions influenced by Na(+) and K(+) binding.

• MeSH
• adenosintrifosfát chemie   MeSH
• červeň bengálská chemie   MeSH
• chemické modely MeSH
• cytoplazma metabolismus   MeSH
• draslík chemie   MeSH
• eosin chemie   MeSH
• erythrosin chemie   MeSH
• Escherichia coli metabolismus   MeSH
• fluorescein chemie   MeSH
• fluoresceiny chemie   farmakologie   MeSH
• fluorescenční barviva farmakologie   MeSH
• konformace proteinů MeSH
• lidé MeSH
• molekulární konformace MeSH
• sodík chemie   MeSH
• sodíko-draslíková ATPasa chemie   MeSH
• terciární struktura proteinů MeSH
• vazebná místa MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

An important mechanism underlying cochlear hair cell (HC) susceptibility to hypoxia/ischemia is the influx of Ca2+. Two main ATP-dependent mechanisms contribute to maintaining low Ca2+ levels: uptake of Ca2+ into intracellular stores via smooth endoplasmic reticulum calcium ATPase (SERCA) and extrusion of Ca2+ via plasma membrane calcium ATPase (PMCA). The effects of the SERCA inhibitors thapsigargin (10 nM-10 µM) and cyclopiazonic acid (CPA; 10-50 µM) and of the PMCA blockers eosin (1.5-10 µM) and o-vanadate (1-5 mM) on inner and outer hair cells (IHCs/OHCs) were examined in normoxia and ischemia using an in vitro model of the newborn rat cochlea. Exposure of the cultures to ischemia resulted in a significant loss of HCs. Thapsigargin and CPA had no effect. Eosin decreased the numbers of IHCs and OHCs by up to 25 % in normoxia and significantly aggravated the ischemia-induced damage to IHCs at 5 and 10 µM and to OHCs at 10 µM. o-Vanadate had no effect on IHC and OHC counts in normoxia, but aggravated the ischemiainduced HC loss in a dose-dependent manner. The effects of eosin and o-vanadate indicate that PMCA has an important role to play in protecting the HCs from ischemic cell death.

• Klíčová slova
• Calcium, Organ of Corti, Ischemia, PMCA, Rat,
• MeSH
• ATPasy přenášející vápník přes plazmatickou membránu antagonisté a inhibitory   MeSH
• Cortiho orgán cytologie   fyziologie   MeSH
• eosin farmakologie   MeSH
• financování organizované MeSH
• inhibitory enzymů farmakologie   MeSH
• ischemie enzymologie   MeSH
• krysa rodu rattus MeSH
• novorozená zvířata MeSH
• perilymfa fyziologie   MeSH
• potkani Wistar MeSH
• sarkoplazmatická Ca2+-ATPáza antagonisté a inhibitory   MeSH
• vanadáty farmakologie   MeSH
• vápník metabolismus   MeSH
• viabilita buněk fyziologie   MeSH
• vnější vláskové buňky účinky léků   MeSH
• vnitřní vláskové buňky účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH

We describe 12 cases of leiomyoma with intracytoplasmic inclusion bodies, which were detected in a group of 447 leiomyomas examined at our institution between December 2005 and March 2006. Ten of these tumors were typical leiomyomas, and two cases represented atypical (bizarre) leiomyoma. In some cases, the presence of intracytoplasmic inclusion bodies resulted in a rhabdoid or skeletal muscle-like appearance of the tumor cells. Ultrastructurally, there were two types of inclusions. One of them consisted of an abnormal aggregation of intermediate and actin filaments. Another type of inclusions was composed of dense granular material without an apparent fibrillar structure. The ultrastructure of the inclusions correlates with immunohistochemical and histochemical stainings. The inclusions with apparent fibrillar arrangements were PAS negative, stained red by trichrome, and were, at least at the periphery, actin-, desmin-, and h-caldesmon-positive. The dense granular inclusions were at least focally PAS-positive, stained red by trichrome, and were negative immunohistochemically. The intracytoplasmic inclusions were found in atypical (bizarre) leiomyomas of the uterus and occasionally in epithelioid leiomyomas and leiomyosarcomas. However, to the best of our knowledge, these inclusions have not been found in typical uterine leiomyomas to date.

• MeSH
• aktiny analýza   MeSH
• azosloučeniny MeSH
• biologické markery analýza   MeSH
• buněčná inkluze chemie   ultrastruktura   MeSH
• desmin analýza   MeSH
• diferenciální diagnóza MeSH
• dospělí MeSH
• eosin MeSH
• financování organizované MeSH
• imunohistochemie metody   MeSH
• keratiny analýza   MeSH
• leiomyom diagnóza   chemie   ultrastruktura   MeSH
• lidé středního věku MeSH
• lidé MeSH
• methylová zeleň MeSH
• MyoD Protein analýza   MeSH
• myogenin analýza   MeSH
• nádory dělohy diagnóza   chemie   ultrastruktura   MeSH
• proteiny vázající kalmodulin analýza   MeSH
• rhabdoidní nádor patologie   MeSH
• Schiffova reakce MeSH
• senioři MeSH
• vimentin analýza   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH

Hearing function in the Fischer 344 (F344) albino inbred strain of rats deteriorates with aging faster than in other strains, in spite of the small hair cell loss in old F344 animals [Popelar, J., Groh, D., Pelanova, J., Canlon, B., Syka, J., 2005. Age-related changes in cochlear and brainstem auditory function. Neurobiol. Aging, in press.]. This study was aimed at elucidating the structural changes in the inner ear of this rat strain during aging. Cochlear histopathology was examined in 20-24-month-old F344 rats and compared with that of young F344 rats (4 months) and of old rats of the Long-Evans (LE) strain. Hematoxylin/eosin staining in aged F344 rats showed degenerative changes in the organ of Corti, consisting of a damaged layer of marginal cells, reduced vascularization of the stria vascularis and a distorted tectorial membrane detached from the organ of Corti. Age-related changes in collagen distribution were observed with Masson's trichrome staining in the spiral ligament of old F344 rats. The results of immunohistochemical staining for type II collagen revealed a marked decrease in collagen fibers in the area connecting the spiral ligament and stria vascularis and a decrease in area IV fibrocytes in old F344 but not in LE rats. These findings may contribute to an explanation of the substantial hearing loss found in old F344 rats.

• MeSH
• azosloučeniny analýza   MeSH
• barvicí látky analýza   MeSH
• Cortiho orgán chemie   MeSH
• eosin analýza   MeSH
• fluorescenční barviva analýza   MeSH
• hematoxylin analýza   MeSH
• imunohistochemie metody   MeSH
• kochlea * fyziologie   chemie   MeSH
• kolagen typ II analýza   MeSH
• kolagen * analýza   MeSH
• krysa rodu rattus MeSH
• membrana tectoria chemie   MeSH
• methylová zeleň analýza   MeSH
• potkani inbrední F344 MeSH
• stárnutí * fyziologie   MeSH
• stria vascularis chemie   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• práce podpořená grantem MeSH

• MeSH
• bronchiální astma diagnóza   farmakoterapie   patologie   MeSH
• bronchoprovokační testy metody   statistika a číselné údaje   využití   MeSH
• eosin MeSH
• eozinofily MeSH
• finanční podpora výzkumu jako téma MeSH
• imunoglobulin E krev   MeSH
• lidé MeSH
• nemoci z povolání diagnóza   etiologie   MeSH
• přehledová literatura jako téma MeSH
• spirometrie MeSH
• sputum chemie   MeSH
• vrcholová výdechová rychlost MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• srovnávací studie MeSH