Myxoinflammatory fibroblastic sarcoma (MIFS) is a rare, low-grade soft tissue neoplasm preferentially arising in the extremities of young to middle-aged adults characterized histologically by a variegated appearance and absence of a distinctive immunophenotype. Herein we have evaluated a series of 73 cases of MIFS to define potential features and markers that may facilitate diagnosis. An immunohistochemical study with a large panel of antibodies showed strong positivity of the tumor cells for bcl-1 (94.5%), FXIIIa (89%), CD10 (80%), and D2-40 (56%). FISH and array comparative genomic hybridization (aCGH) were performed in a large subset of cases to investigate the utility for detecting the TGFBR3 and OGA t(1;10) rearrangement and BRAF abnormalities. Using a combination of FISH and/or aCGH, t(1;10) was detected in only 3 of 54 cases (5.5%). The aCGH study also demonstrated amplification of VGLL3 on chromosome 3 that was detected in 8 of 20 cases (40%). BRAF alterations were observed by FISH in 4 of 70 cases (5.7%) and correlated with gain of chromosome 3p12 (VGLL3). A novel fusion transcript involving exon 6 of ZNF335 and exon 10 of BRAF was identified in one case. Demonstration of amplification of VGLL3 on chromosome 3 in combination with expression of bcl-1 and FXIIIa may help support the diagnosis, however, due to their low specificity these markers are not sufficient for a definitive diagnosis in the absence of the appropriate clinical-pathological context. Until a more robust genetic or immunohistochemical signature is identified, the diagnosis of MIFS rests on its characteristic clinicopathological features.
- MeSH
- amplifikace genu MeSH
- diagnostické techniky molekulární * MeSH
- dospělí MeSH
- fenotyp MeSH
- fibroblasty chemie patologie MeSH
- fibrosarkom chemie genetika patologie MeSH
- fúze genů MeSH
- genetická predispozice k nemoci MeSH
- genová přestavba MeSH
- hybridizace in situ fluorescenční MeSH
- imunohistochemie * MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- nádorové biomarkery * analýza genetika MeSH
- nádory měkkých tkání chemie genetika patologie MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- srovnávací genomová hybridizace MeSH
- translokace genetická MeSH
- vysoce účinné nukleotidové sekvenování MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- senioři nad 80 let MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- multicentrická studie MeSH
- Geografické názvy
- Evropa MeSH
- Spojené státy americké MeSH
Singlet oxygen is a highly reactive species which is involved in a number of processes, including photodynamic therapy of cancer. Its very weak near-infrared emission makes imaging of singlet oxygen in biological systems a long-term challenge. We address this challenge by introducing Singlet Oxygen Feedback Delayed Fluorescence (SOFDF) as a novel modality for semi-direct microscopic time-resolved wide-field imaging of singlet oxygen in biological systems. SOFDF has been investigated in individual fibroblast cells incubated with a well-known photosensitizer aluminium phthalocyanine tetrasulfonate. The SOFDF emission from the cells is several orders of magnitude stronger and much more readily detectable than the very weak near-infrared phosphorescence of singlet oxygen. Moreover, the analysis of SOFDF kinetics enables us to estimate the lifetimes of the involved excited states. Real-time SOFDF images with micrometer spatial resolution and submicrosecond temporal-resolution have been recorded. Interestingly, a steep decrease in the SOFDF intensity after the photodynamically induced release of a photosensitizer from lysosomes has been demonstrated. This effect could be potentially employed as a valuable diagnostic tool for monitoring and dosimetry in photodynamic therapy.
- MeSH
- buňky 3T3 MeSH
- časové faktory MeSH
- fibroblasty chemie cytologie MeSH
- fluorescence * MeSH
- fluorescenční mikroskopie MeSH
- fotochemoterapie MeSH
- fotosenzibilizující látky chemie MeSH
- indoly chemie MeSH
- kultivované buňky MeSH
- myši MeSH
- optické zobrazování * MeSH
- organokovové sloučeniny chemie MeSH
- singletový kyslík analýza chemie MeSH
- viabilita buněk MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
In this study, biodegradable poly(ε-caprolactone) (PCL) nanofibers (PCL-NF), collagen-coated PCL nanofibers (Col-c-PCL), and titanium dioxide-incorporated PCL (TiO2-i-PCL) nanofibers were prepared by electrospinning technique to study the surface and structural compatibility of these scaffolds for skin tisuue engineering. Collagen coating over the PCL nanofibers was done by electrospinning process. Morphology of PCL nanofibers in electrospinning was investigated at different voltages and at different concentrations of PCL. The morphology, interaction between different materials, surface property, and presence of TiO2 were studied by scanning electron microscopy (SEM), Fourier transform IR spectroscopy (FTIR), contact angle measurement, energy dispersion X-ray spectroscopy (EDX), and X-ray photoelectron spectroscopy (XPS). MTT assay and cell adhesion study were done to check biocompatibilty of these scaffolds. SEM study confirmed the formation of nanofibers without beads. FTIR proved presence of collagen on PCL scaffold, and contact angle study showed increment of hydrophilicity of Col-c-PCL and TiO2-i-PCL due to collagen coating and incorporation of TiO2, respectively. EDX and XPS studies revealed distribution of entrapped TiO2 at molecular level. MTT assay and cell adhesion study using L929 fibroblast cell line proved viability of cells with attachment of fibroblasts over the scaffold. Thus, in a nutshell, we can conclude from the outcomes of our investigational works that such composite can be considered as a tissue engineered construct for skin wound healing.
- MeSH
- biokompatibilní materiály chemie MeSH
- buněčná adheze fyziologie MeSH
- fibroblasty chemie MeSH
- fotoelektronová spektroskopie metody MeSH
- hydrofobní a hydrofilní interakce MeSH
- kolagen chemie MeSH
- kůže chemie MeSH
- nanovlákna chemie MeSH
- polyestery chemie MeSH
- povrchové vlastnosti MeSH
- titan chemie MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury MeSH
- Publikační typ
- časopisecké články MeSH
The present work provides a proof-of-concept that the singlet oxygen-sensitized delayed fluorescence (SOSDF) can be detected from individual living mammalian cells in a time-resolved microscopy experiment. To this end, 3T3 mouse fibroblasts incubated with 100 μM TPPS4 or TMPyP were used and the microsecond kinetics of the delayed fluorescence (DF) were recorded. The analysis revealed that SOSDF is the major component of the overall DF signal. The microscopy approach enables precise control of experimental conditions - the DF kinetics are clearly influenced by the presence of the (1)O2 quencher (sodium azide), H2O/D2O exchange, and the oxygen concentration. Analysis of SOSDF kinetics, which was reconstructed as a difference DF kinetics between the unquenched and the NaN3-quenched samples, provides a cellular (1)O2 lifetime of τΔ = 1-2 μs and a TPPS4 triplet lifetime of τT = 22 ± 5 μs in agreement with previously published values. The short SOSDF acquisition times, typically in the range of tens of seconds, enable us to study the dynamic cellular processes. It is shown that SOSDF lifetimes increase during PDT-like treatment, which may provide valuable information about changes of the intracellular microenvironment. SOSDF is proposed and evaluated as an alternative tool for (1)O2 detection in biological systems.
- MeSH
- analýza jednotlivých buněk přístrojové vybavení metody MeSH
- azid sodný chemie MeSH
- buňky 3T3 MeSH
- design vybavení MeSH
- fibroblasty chemie MeSH
- fluorescence * MeSH
- kinetika MeSH
- kyslík chemie MeSH
- mikroskopie přístrojové vybavení metody MeSH
- myši MeSH
- oxid deuteria chemie MeSH
- singletový kyslík chemie MeSH
- voda chemie MeSH
- zvířata MeSH
- Check Tag
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
F1Fo-ATP synthase is a key enzyme of mitochondrial energy provision producing most of cellular ATP. So far, mitochondrial diseases caused by isolated disorders of the ATP synthase have been shown to result from mutations in mtDNA genes for the subunits ATP6 and ATP8 or in nuclear genes encoding the biogenesis factors TMEM70 and ATPAF2. Here, we describe a patient with a homozygous p.Tyr12Cys mutation in the epsilon subunit encoded by the nuclear gene ATP5E. The 22-year-old woman presented with neonatal onset, lactic acidosis, 3-methylglutaconic aciduria, mild mental retardation and developed peripheral neuropathy. Patient fibroblasts showed 60-70% decrease in both oligomycin-sensitive ATPase activity and mitochondrial ATP synthesis. The mitochondrial content of the ATP synthase complex was equally reduced, but its size was normal and it contained the mutated epsilon subunit. A similar reduction was found in all investigated F1 and Fo subunits with the exception of Fo subunit c, which was found to accumulate in a detergent-insoluble form. This is the first case of a mitochondrial disease due to a mutation in a nuclear encoded structural subunit of the ATP synthase. Our results indicate an essential role of the epsilon subunit in the biosynthesis and assembly of the F1 part of the ATP synthase. Furthermore, the epsilon subunit seems to be involved in the incorporation of subunit c to the rotor structure of the mammalian enzyme.
- MeSH
- fibroblasty enzymologie chemie MeSH
- kultivované buňky MeSH
- lidé MeSH
- missense mutace * MeSH
- mitochondriální nemoci * enzymologie genetika MeSH
- mitochondriální protonové ATPasy * genetika nedostatek MeSH
- mladý dospělý MeSH
- molekulární sekvence - údaje MeSH
- proteiny genetika chemie metabolismus MeSH
- sekvence aminokyselin MeSH
- sekvence nukleotidů MeSH
- sekvenční seřazení MeSH
- Check Tag
- lidé MeSH
- mladý dospělý MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
The transplacental cell transfer naturally takes place during pregnancy and occurs bi-directionally between the mother and fetus. Using real-time polymerase chain reaction (PCR) assay and sex determining region Y (SRY) gene as a marker, we examined the presence of male fetal cells in cell cultures derived from synovial tissues and skin dermis in women with prior pregnancy history suffering from rheumatoid arthritis (RA) who underwent synovectomy. Male DNA was detected in synovial cell samples derived from carpal, hip, metacarpophalangeal and metatarsophalangeal joints in five out of 13 (38.5%) patients with RA in a frequency range of 0.02-62.55 (mean 12.17) male cells per 10,000,000 total cells. SRY gene positivity was found as well in skin fibroblast cultures in four out of 10 (40.0%) RA patients in a frequency range of 3.26-43.47 (mean 15.42) male cells per 10,000,000 total cells, respectively. The difference in a frequency of fetal-derived male cells between both the cohorts did not achieve the statistical difference (p=0.77). We conclude that persisting male fetal cells are able to grow from non-inflamed tissues as well as from those which have many features characteristic of a stressed tissue. We conclude that persisting male fetal cells are also able to proliferate in cell culture since their presence was detected even in consecutive passages.
- MeSH
- chimérismus MeSH
- DNA analýza MeSH
- dospělí MeSH
- fibroblasty chemie MeSH
- genetické markery genetika MeSH
- kultivované buňky MeSH
- kůže cytologie MeSH
- lidé MeSH
- mezoderm cytologie růst a vývoj MeSH
- mladý dospělý MeSH
- plod cytologie MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- protein oblasti určující pohlaví na chromozomu Y genetika MeSH
- revmatoidní artritida komplikace MeSH
- synovektomie MeSH
- synoviální membrána cytologie chemie MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- mladý dospělý MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- práce podpořená grantem MeSH
Mitochondrial dysfunction of the energy generating system was suggested in two infants with progressive infantile poliodystrophy characterised by hypotonia, refractory epilepsy, visual impairment, psychomotor retardation, profound brain atrophy, hepatopathy, and increased levels of lactate in blood and cerebrospinal fluid. Histochemical and electron microscopic analyses of liver biopsies revealed cytochrome c oxidase deficiency, microvesicular steatosis, and enormous multiplication of mitochondria of various sizes. In the first patient, the quantitative Southern blot analyses in tissues obtained at autopsy demonstrated reduced content of mtDNA in the liver, brain, and fibroblasts (11 %, 15 %, and 25 % of the mean values in controls) while a normal content of mtDNA was found in muscle and heart. In the second patient, a reduced content of mtDNA was found in the muscle, liver, and brain (15 %, 10 %, and 30 %, respectively, of the mean values in controls). Biochemical studies in the first patient revealed decreased activities of all respiratory chain complexes except complex II in isolated liver mitochondria and decreased amounts of respiratory chain complexes I, III, IV and ATP synthase in liver and frontal cortex, but not in muscle, heart, and fibroblasts. In conclusions, mtDNA depletion associated with Alpers syndrome may be tissue specific.
- MeSH
- fibroblasty chemie MeSH
- játra chemie patologie MeSH
- kojenec MeSH
- lidé MeSH
- mitochondriální DNA * analýza genetika MeSH
- mozek - chemie MeSH
- mozek patologie MeSH
- Schilderova difuzní cerebroskleróza * genetika metabolismus patologie MeSH
- svaly chemie patologie MeSH
- Check Tag
- kojenec MeSH
- lidé MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
- MeSH
- experimentální sarkom MeSH
- fibroblasty fyziologie chemie MeSH
- finanční podpora výzkumu jako téma MeSH
- kalibrace MeSH
- krysa rodu rattus MeSH
- kultivované buňky MeSH
- listy rostlin cytologie fyziologie MeSH
- pohyb buněk MeSH
- zobrazování trojrozměrné MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- buněčné jádro účinky záření MeSH
- fibroblasty chemie účinky záření ultrastruktura MeSH
- filadelfský chromozom MeSH
- geny abl MeSH
- lidé MeSH
- lidské chromozomy, pár 22 genetika účinky záření ultrastruktura MeSH
- lidské chromozomy, pár 9 genetika účinky záření ultrastruktura MeSH
- lymfocyty ultrasonografie MeSH
- Check Tag
- lidé MeSH
