Analysis of epidermal genes, proteins and lipids is important in the research and diagnosis of skin diseases. Although punch biopsy is the first-choice technique for the skin sampling, it is unnecessarily invasive for obtaining a sample just for the epidermal analysis. Here we compare two less invasive methods, suction blistering (SB) and tape stripping (TS), for the analysis of selected epidermal genes (quantitative real-time reverse transcription PCR, qRT-PCR), proteins (western blotting, WB), and lipids in ten healthy volunteers. TS provided significantly less material than SB and no viable epidermal layers could be obtained according to the reflectance confocal microscopy. Consistently, only the SC protein filaggrin and housekeeping GAPDH together with FLG and RPL13A mRNA were detected by TS. In the SB samples, WB and qRT-PCR could easily detect all the selected proteins (claudin-1, occludin, filaggrin, laminin and GAPDH) and genes (CLDN1, OCLN, FLG, LAMA3 and RPL13A), respectively. A single SB sample further provided enough of material for immunohistochemistry and lipid analyses, which was not feasible with the TS samples. Immunohistochemistry of the SB samples showed intact epidermal structure and a characteristic expression of claudin-1. Infrared spectroscopy showed well-ordered lipids with both orthorhombic and hexagonal packing and high-performance thin layer chromatography confirmed all lipid classes (including ceramide subclasses) in correct proportions. Taken together, SB represents a reliable sampling technique that can be utilized for multipurpose epidermal analyses in various studies.

• MeSH
• chromatografie na tenké vrstvě MeSH
• claudin-1 analýza   MeSH
• dospělí MeSH
• epidermis chemie   MeSH
• imunohistochemie MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipidy analýza   MeSH
• messenger RNA analýza   MeSH
• odsávání MeSH
• proteiny intermediálních filament analýza   MeSH
• proteiny analýza   MeSH
• puchýř MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Identifikace nádorových kmenových buněk v současnosti představuje jednu z nejdůležitějších oblastí výzkumu, neboť nádorové kmenové buňky hrají důležitou úlohu v iniciaci a progresi nádoru, stejně jako v procesech metastazování a relapsu onemocnění. Tento článek shrnuje současné poznatky o známých i předpokládaných markerech nádorových kmenových buněk v různých typech sarkomů kostí i měkkých tkání. Zvláštní pozornost je věnována detekci CD133, ABC transportérů, nestinu a aldehyddehydrogenázy, které byly v posledních letech intenzivně zkoumány jak v nádorové tkáni, tak v sarkomových buněčných liniích. V závěru článku je uveden přehled možných fenotypů nádorových kmenových buněk, které byly prokázány funkčními testy tumorigenicity.

The identification of cancer stem cell markers represents one of the very relevant research to­pics because cancer stem cells play important roles in tumour initiation and progression, as well as during metastasis formation and in relapse of the disease. This article summarises recent knowledge on well-known and putative cancer stem cell markers in various types of bone and soft-tissue sarcomas. Special attention is paid to the detection of CD133, ABC transporters, nestin and aldehyde dehydrogenase that have been intensively studied both in tumour tissues and in sarcoma cell lines during the past few years. Finally, an overview is given of the possible CSC phenotypes provided by functional assays of tumourigenicity.

• Klíčová slova
• tumorigenicita, aldehyddehydrogenáza, CD133, rabdomyosarkom,
• MeSH
• ABC transportéry analýza   genetika   MeSH
• aldehyddehydrogenasa analýza   genetika   MeSH
• CD antigeny analýza   genetika   MeSH
• glykoproteiny analýza   genetika   MeSH
• lidé MeSH
• nádorové biomarkery analýza   MeSH
• nádorové buněčné linie MeSH
• nádorové kmenové buňky * patologie   MeSH
• nestin MeSH
• osteosarkom diagnóza   genetika   MeSH
• peptidy analýza   genetika   MeSH
• proteiny intermediálních filament analýza   genetika   MeSH
• rhabdomyosarkom diagnóza   genetika   MeSH
• sarkom * diagnóza   genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH

Nestin, marker multipotentních prekurzorových buněk, představuje významnou dynamickou strukturu, jejíž polymerizace/depolymerizace ovlivňuje intracelulární signalizaci a podílí se na řadě klíčových buněčných procesů, jako je proliferace, migrace a přežívání buněk. Předpokládá se, že nestin hraje centrální roli v procesu karcinogeneze. Nestin je považován za možný diagnostický a prediktivní indikátor malignity solidních nádorů a potenciální marker nádorových kmenových buněk. Překvapivě byl identifikován i ve zralých CD138+38+ plazmatických buňkách (PC) mnohočetného myelomu (MM). Exprese markeru kmenových/progenitorových buněk v maligních PC, které jsou považovány za terminálně diferencované, indikuje, že nestin by mohl hrát významnou roli v patologii MM.

Nestin, a marker of multipotent precursor cells, is an important dynamic structure; its polymerization/ depolymerization influences intracellular signaling and participates in key cell processes such as proliferation, migration and cell survival. It is presumed that nestin plays a central role in carcinogenesis. It is suggested that nestin might be a suitable diagnostic and prognostic indicator of malignancy and a potential marker of cancer stem cells. Unexpectedly, nestin has been identified in mature CD138+CD38+ plasma cells (PC) of multiple myeloma patients (MM). Expression of nestin, a marker of stem/progenitor cells, in malignant PC, that are considered to be terminally differentiated, indicates that nestin might play a unique role in pathology of MM.

• Klíčová slova
• flowcytometrie, iniciující buňky, myelomové kmenové buňky,
• MeSH
• financování organizované MeSH
• lidé MeSH
• mnohočetný myelom diagnóza   metabolismus   patofyziologie   MeSH
• nádorové biomarkery MeSH
• plazmatické buňky MeSH
• proteiny intermediálních filament analýza   fyziologie   MeSH
• proteiny nervové tkáně analýza   fyziologie   MeSH
• Check Tag
• lidé MeSH

BACKGROUND AND AIMS: Nestin is considered to be a marker of stem/progenitor cells in different tissues. Nestin expression was also described in various tumors. In pancreatic ductal adenocarcinoma (PDAC), its role in cancer cell migration, invasion, and metastases has been suggested. The study aimed at examining the expression of nestin in PDAC, and to evaluate its clinicopathological correlations. METHODS: The expression of nestin was immunohistochemically examined in 117 PDAC resection specimens, analyzed, and correlated with clinico-pathological parameters including perineural invasion (PNI). Analysis of nestin expression in nerve fibers in tissues of chronic pancreatitis (CP) was added. RESULTS: Immunohistochemical analysis of nestin expression showed 79 nestin negative (67.5 %) and 38 nestin positive (32.5 %) PDACs. No significant correlations of nestin expression in tumor cells with the analyzed clinicopathological parameters were demonstrated. Tumor grade (p<0.001) and nodal status (p=0.009) proved to represent independent prognostic factors. PNI was identified in 94 PDAC (80.3 %), and did not correlate with nestin expression. Nestin immunostaining was displayed in nerve fibers of both CP and PDAC tissues. CONCLUSION: An intimate link of nestin to a biological process of pancreatic cancer was confirmed. The expression of nestin did not prove to be a valuable prognostic factor and an immunohistochemical assessment of nestin expression is not superior to conventional prognostic factors in PDAC. A correlation between nestin expression in tumor cells and PNI was not confirmed and expression of nestin in nerve fibers of both PDAC and CP tissues seems to reflect the process of neural remodeling responsible for pancreatic neuropathy.

• MeSH
• duktální karcinom pankreatu chemie   mortalita   sekundární   chirurgie   MeSH
• imunohistochemie MeSH
• invazivní růst nádoru MeSH
• Kaplanův-Meierův odhad MeSH
• lidé středního věku MeSH
• lidé MeSH
• lymfatické metastázy MeSH
• nádorové biomarkery analýza   MeSH
• nádory slinivky břišní chemie   mortalita   patologie   chirurgie   MeSH
• nervová vlákna patologie   MeSH
• pankreatektomie MeSH
• prediktivní hodnota testů MeSH
• prognóza MeSH
• proteiny intermediálních filament analýza   MeSH
• proteiny nervové tkáně analýza   MeSH
• rozdělení chí kvadrát MeSH
• senioři MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

Nestin is a class VI intermediate filament protein expressed in the cytoplasm of stem and progenitor cells in the mammalian CNS during development. In adults, nestin is present only in a small subset of cells and tissues, including the subventricular zone of the adult mammalian brain, where neurogenesis occurs. Nestin expression has also been detected under such pathological conditions as ischemia, inflammation, and brain injury, as well as in various types of human solid tumors and their corresponding cell lines. Furthermore, nestin was recently found in the nuclei of glioblastoma, neuroblastoma, and angiosarcoma cells and it was proved to interact directly with the nuclear DNA in neuroblastoma cells. Here, we perform the first study of the intracellular distribution of nestin in cell lines derived from neurogenic tumors. Using immunodetection methods, we examined nestin expression in tumor-derived cell lines obtained from 11 patients with neuroblastoma, medulloblastoma, or glioblastoma multiforme. Besides its standard cytoplasmic localization, nestin was present in the nuclei of two neuroblastoma cell lines and one medulloblastoma cell line. Nestin was only present in the nuclei of cells with diffuse cytoplasmic staining for this protein, and the proportion of cells positive for nestin in nuclei, as well as the intensity of staining, varied. The presence of nestin in the nuclei was confirmed by both transmission electron microscopy and Western blotting. Our results indicate that the presence of nestin in the nuclei of tumor cells is not very rare, especially under in vitro conditions.

• MeSH
• buněčné jádro chemie   metabolismus   ultrastruktura   MeSH
• dítě MeSH
• fluorescenční protilátková technika MeSH
• glioblastom metabolismus   ultrastruktura   MeSH
• imunohistochemie MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• meduloblastom metabolismus   ultrastruktura   MeSH
• nádorové buněčné linie MeSH
• neuroblastom metabolismus   ultrastruktura   MeSH
• předškolní dítě MeSH
• proteiny intermediálních filament analýza   metabolismus   MeSH
• proteiny nervové tkáně analýza   metabolismus   MeSH
• senioři MeSH
• transmisní elektronová mikroskopie MeSH
• western blotting MeSH
• Check Tag
• dítě MeSH
• kojenec MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

AIM: To describe the case history and new histopathological findings of a young woman suffering from moyamoya disease. METHODS: The patient underwent brain computed tomography, magnetic resonance imaging and brain angiography. Vessel samples of a. temporalis superficialis were processed by standard histopathological and immunohistochemical methods by analysis of VEGF, VEGFR and nestin expression. RESULTS: Brain angiography revealed both internal carotid artery stenoses and stenoses of the anterior cerebral arteries. Stenotic parts of vessels were accompanied by coiled and elongated vessels with a picture of "smoke puffs carried away by breeze" after contrast medium application. Histological examination showed: obstruction of lumen, fibrocellular intimal thickening, tortuosity and disruption of internal elastic lamina. Imunohistochemistry confirmed a defect of the internal elastic membrane of the muscular arteria and progressive intimal thickening accompanied by abnormal smooth muscle cells and, VEGF/VEGFR expression in intima. Nestin positivity in endothelium of arteria indicated that endothelial cells are activated. CONCLUSION: We found that the endothelium of affected vessels is nestin positive. This, together with the finding of VEGF/VEGFR expression, might suggest an active angiogenetic process We present a new conception of pathogenesis but further studies with higher number of patients are necessary to elucidate the role of these growth factors in the moyamoya disease.

• MeSH
• arteriae carotides radiografie   MeSH
• cévní endotel metabolismus   patofyziologie   patologie   MeSH
• dospělí MeSH
• financování organizované MeSH
• imunohistochemie MeSH
• lidé MeSH
• moyamoya nemoc MeSH
• proteiny intermediálních filament analýza   MeSH
• proteiny nervové tkáně analýza   MeSH
• receptory vaskulárního endoteliálního růstového faktoru analýza   MeSH
• vaskulární endoteliální růstové faktory analýza   MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH

We describe the expression and distribution patterns of nestin, desmin and vimentin in intact and regenerating muscle spindles of the rat hind limb skeletal muscles. Regeneration was induced by intramuscular isotransplantation of extensor digitorum longus (EDL) or soleus muscles from 15-day-old rats into the EDL muscle of adult female inbred Lewis rats. The host muscles with grafts were excised after 7-, 16-, 21- and 29-day survival and immunohistochemically stained. Nestin expression in intact spindles in host muscles was restricted to Schwann cells of sensory and motor nerves. In transplanted muscles, however, nestin expression was also found in regenerating "spindle fibers", 7 and 16 days after grafting. From the 21st day onwards, the regenerated spindle fibers were devoid of nestin immunoreactivity. Desmin was detected in spindle fibers at all developmental stages in regenerating as well as in intact spindles. Vimentin was expressed in cells of the outer and inner capsules of all muscle spindles and in newly formed myoblasts and myotubes of regenerating spindles 7 days after grafting. Our results show that the expression pattern of these intermediate filaments in regenerating spindle fibers corresponds to that found in regenerating extrafusal fibers, which supports our earlier suggestion that they resemble small-diameter extrafusal fibers.

• MeSH
• desmin analýza   MeSH
• financování organizované MeSH
• kosterní svalová vlákna chemie   MeSH
• kosterní svaly chemie   transplantace   ultrastruktura   MeSH
• krysa rodu rattus MeSH
• myoblasty chemie   MeSH
• nervosvalová vřeténka chemie   MeSH
• potkani inbrední LEW MeSH
• proteiny intermediálních filament analýza   MeSH
• proteiny nervové tkáně analýza   MeSH
• regenerace MeSH
• Schwannovy buňky chemie   MeSH
• vimentin analýza   MeSH
• zadní končetina MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• ženské pohlaví MeSH
• zvířata MeSH

• MeSH
• fyziologie kůže MeSH
• homeostáza fyziologie   MeSH
• mutantní kmeny myší MeSH
• proteiny intermediálních filament analýza   klasifikace   ultrastruktura   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH

Effects of active ingredients from topical and systemic skincare products on structure and organization of epidermis, dermal-epidermal junction (DEJ), and dermis were examined using an in vitro reconstructed skin equivalent (SE). Imedeen Time Perfection (ITP) ingredients (a mixture of BioMarine Complex, grape seed extract, tomato extract, vitamin C) were supplemented systemically into culture medium. Kinetin, an active ingredient from Imedeen Expression Line Control Serum, was applied topically. Both treatments were tested separately or combined. In epidermis, all treatments stimulated keratinocyte proliferation, showing a significant increase of Ki67-positive keratinocytes (P < 0.05). Kinetin showed a twofold increase of Ki67-positive cells, ITP resulted in a fivefold, and ITP+kinetin showed a nine-fold increase. Differentiation of keratinocytes was influenced only by kinetin since filaggrin was found only in kinetin and kinetin+ITP samples. At the DEJ, laminin 5 was slightly increased by all treatments. In dermis, only ITP increased the amount of collagen type I. Both kinetin and ITP stimulated formation of fibrillin-1 and elastin deposition. The effect of kinetin was seen in upper dermis. It stimulated not only the amount of deposited fibrillin-1 and elastin fibers but also their organization perpendicularly to the DEJ. ITP stimulated formation of fibrillin-1 in deeper dermis. In summary, the combination of topical treatment with kinetin and systemic treatment with ITP had complementary beneficial effects in the formation and development of epidermis and dermis.

• MeSH
• antigen Ki-67 metabolismus   MeSH
• aplikace lokální MeSH
• buněčné kultury MeSH
• dospělí MeSH
• elastin metabolismus   MeSH
• epidermis cytologie   MeSH
• glykosaminoglykany aplikace a dávkování   farmakologie   MeSH
• keratinocyty cytologie   metabolismus   MeSH
• kinetin metabolismus   MeSH
• kultivované buňky MeSH
• kůže MeSH
• lékové interakce MeSH
• lidé MeSH
• mikrofilamentové proteiny metabolismus   MeSH
• péče o kůži MeSH
• proteiny intermediálních filament analýza   metabolismus   MeSH
• proteiny aplikace a dávkování   farmakologie   MeSH
• regulátory růstu rostlin farmakologie   MeSH
• škára cytologie   MeSH
• umělá kůže MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• srovnávací studie MeSH

BACKGROUND: Nestin is a protein belonging to class VI of intermediate filaments that is produced in stem/progenitor cells in the mammalian CNS during development and is consecutively replaced by other intermediate filament proteins (neurofilaments, GFAP). Down-regulated nestin may be re-expressed in the adult organism under certain pathological conditions (brain injury, ischemia, inflammation, neoplastic transformation). Our work focused on a detailed study of the nestin cytoskeleton in cell lines derived from glioblastoma multiforme, because re-expression of nestin together with down-regulation of GFAP has been previously reported in this type of brain tumor. METHODS: Two cell lines were derived from the tumor tissue of patients treated for glioblastoma multiforme. Nestin and other cytoskeletal proteins were visualized using imunocytochemical methods: indirect immunofluorescence and immunogold-labelling. RESULTS: Using epifluorescence and confocal microscopy, we described the morphology of nestin-positive intermediate filaments in glioblastoma cells of both primary cultures and the derived cell lines, as well as the reorganization of nestin during mitosis. Our most important result came through transmission electron microscopy and provided clear evidence that nestin is present in the cell nucleus. CONCLUSION: Detailed information concerning the pattern of the nestin cytoskeleton in glioblastoma cell lines and especially the demonstration of nestin in the nucleus represent an important background for further studies of nestin re-expression in relationship to tumor malignancy and invasive potential.

• MeSH
• cytoskelet chemie   ultrastruktura   MeSH
• financování organizované MeSH
• fluorescenční protilátková technika nepřímá MeSH
• glioblastom genetika   chemie   patologie   ultrastruktura   MeSH
• gliový fibrilární kyselý protein analýza   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• proteiny intermediálních filament analýza   MeSH
• proteiny nervové tkáně analýza   MeSH
• vimentin analýza   MeSH
• Check Tag
• lidé MeSH