K detekci apoptózy (AP) na tkáňových řezech lze využít morfologické hodnocení AP figur světelnou či elektronovou mikroskopií a imunohistochemícky průkaz antigénu asociovaných s AP. Bcl2 u řady nádorů inverzně koreluje s AP a je nepřímým markerem počtu AP figur. Imunohistochemícky průkaz je bezproblémový. Tkáňovou transglutaminázu, která je výrazně exprimována i v neapoptotických buňkách, nelze považovat za specifický marker AP. Podobně FAS, jehož exprese nemusí být bezpodmínečně spojena se vstupem buňky do AP. Kaspázy: v současnosti používaná imunohistochemická detekce nerozlišuje aktivní a neaktivní formy těchto enzymů. Anexin V: technika, detekující fosfatidylserin, normálně lokalizovaný na vnitřní straně buněčné membrány, jenž je při apoptotickém procesu detekovatelný i extracelulámě. Průkaz Anexinu V značeného koloidním zlatem lze využít k detekci AP imunoelektronovou mikroskopií, nicméně jde o velmi náročnou metodiku. TUNEL, ISEL, ISNTA jsou metody, které označují zlomy DNA vznikající při apoptóze. TUNEL ve srovnání s morfologickým hodnocením AP figur detekuje o cca 20 % apoptotických buněk více. Označeny mohou být i fragmenty DNA vznikající v jádrech během reparace, po elektrokoagulaci, při autolýze, následkem fixace a řezání tkání. Apostain je metoda založená na průkazu jednovláknových zlomů DNA pomocí monoklonální protilátky, která využívá sníženou stabilitu apoptotické DNA vůči termální denaturaci způsobující její zlomy. Protilátka M30 CytoDeath detekuje neoepitopy vzniklé při štěpení cytokeratinů 18 kaspázami. Ve srovnání s TUNELem chybí exprese v nekrotických buňkách, neoznačuje však buňky v pokročilých stadiích AP (tělíska). Na rozdíl od některých pilotních studií se nám u M30 (Roche) nepodařilo docílit dostatečnou senzitivitu ani specificitu při detekci AP na mamámích karcinomech. Rutinní, resp. automatizované hodnocení AP vyžaduje ověření a standardizaci dalších metodik.

Signs Methods for identification of apoptotic (AP) cells in tissue sections include light and electron microscopy and immunohistochemical (IHC) detection of apoptotic antigens. BcI-2 at many tumors inversely correlates with AP and is an indirect marker of AP index. Transglutaminase is often expressed in nonapoptotic cells, and thus represents a non specific marker of AP. Likewise, expression of FAS does not necessarily represent a transformation into AP. IHC detection of caspases does not distinguish between active and nonactive forms of the proteases. Immunolabeling of biotin-conjugated Annexin V is used for the identification of phosphatidylserine residues exposed on the surface of AP cells. Annexin V immuno-gold labeling by means of electron microscopy will allow a more refined description of the morphological events occurring during apoptosis. TUNEL, ISEL and ISNTA methods detect DNA breaks. The rate of AP detected by TUNEL is about 20 % higher then by apoptotic figure counting. DNA strand breaks can also occur during DNA repair, electrocoagulation, autolysis, fixation and paraffin embedding. With Apostain, DNA is selectively denaturated bv heatinef with formamide and stained by monoclonal antibody specific to single-strand DNA. It specifically stains condensed chromatin of apoptotic cells. M30 IHC uses a monoclonal antibody binding to the product resulting from cleavage of cytokeratin 18 by activated caspases. M30 is negative in necrotic cells and in progressively degraded cells (AP bodies). In contrast to some pilot studies, we have not reached sufficient sensitivity and specificity of IHC detection with M30 (Roche) in breast carcinomas.

• MeSH
• antigeny nádorové analýza   MeSH
• apoptóza genetika   MeSH
• biologické markery analýza   MeSH
• buněčná smrt MeSH
• elektronová mikroskopie metody   MeSH
• finanční podpora výzkumu jako téma MeSH
• imunohistochemie metody   MeSH
• nádorové kmenové buňky MeSH
• techniky in vitro MeSH

• MeSH
• databáze jako téma MeSH
• histologie * výchova   MeSH
• informační systémy MeSH
• počítačem řízená výuka MeSH
• programovaná výuka jako téma * MeSH
• studium lékařství metody   MeSH
• výuka - hodnocení metody   MeSH
• využití lékařské informatiky MeSH
• Geografické názvy
• Česká republika MeSH

• Publikační typ
• abstrakt z konference MeSH

• MeSH
• histologické techniky MeSH
• histologie MeSH

• Konspekt
• Anatomie člověka a srovnávací anatomie
• NLK Obory
• histologie
• NLK Publikační typ
• praktická cvičení
• učebnice vysokých škol

• MeSH
• histologické techniky MeSH

• Konspekt
• Buněčná biologie. Cytologie
• NLK Obory
• cytologie, klinická cytologie
• histologie
• NLK Publikační typ
• učebnice vysokých škol

INTRODUCTION: Virtual microscopy, used as a method to teach histology, has many undeniable advantages. However, the usefulness of this method is somewhat limited by the difficulties students face in finding their way through huge amounts of digital data, compounded by decreased interaction between students and teachers. We describe the results of a recent pilot project which combined the modern teaching methods of active learning, where students themselves present histological slides and make use of the virtual microscopy system. METHODS: Students' responses to a structured questionnaire and examination results were evaluated. RESULTS: We found that a combination of both electronic materials and textbooks was commonly used by students to prepare for practical teaching sessions, with electronic resources being used regularly by the majority of students. No statistically relevant differences were found between the approaches of dentistry vs general medicine students. Cooperation between students' groups during the preparation for individual presentations was seen to be beneficial by a majority of dentistry students; they reported that the introduction of student-led presentations improved their quality of preparation for practical lessons, as well as increasing their participation and activity level in the lessons themselves. These different approaches and motivations between students of dentistry and general medicine are reflected in the test results where dentistry students are more successful. CONCLUSION: We confirm that there are differences in motivation, approaches and examination results between both groups of students, which should be taken into account and which could lead to differentiation of future curricula for both study courses.

• MeSH
• histologie výchova   MeSH
• lidé MeSH
• mikroskopie metody   MeSH
• pilotní projekty MeSH
• průzkumy a dotazníky MeSH
• studenti lékařství psychologie   MeSH
• studenti stomatologie psychologie   MeSH
• studium stomatologie metody   MeSH
• učební pomůcky MeSH
• učení * MeSH
• virtuální realita * MeSH
• vyučování * MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

• MeSH
• histologické techniky MeSH
• histologie MeSH

• Konspekt
• Výchova a vzdělávání
• NLK Obory
• zdravotní výchova
• histologie
• NLK Publikační typ
• učebnice vysokých škol

BACKGROUND: The generally accepted indications for carotid endarterectomy are the clinical picture and degree of per cent stenosis of the carotid artery. Despite the fact that stenosis measurement is defined, the methods vary considerably. The correlation of particular methods, especially angiography and duplex sonography, has been repeatedly demonstrated. However, the correlation between any technique and true anatomical stenosis, as evaluated on the surgical specimen, has been only anecdotally reported. METHOD: During carotid endarterectomy, the atherosclerotic plaque was removed in one piece and subsequently stored and histologically processed. The histological slides were evaluated under an optical microscope, scanned and the slide with maximum stenosis was determined using a planimetric program. Both the minimal lumen area and the area of the whole plaque were measured. The stenosis was calculated using the planimetric method. On the maximum stenosis slice, the minimal diameter and the diameter of the whole plaque were also measured. Angiographic images were scanned and the per cent stenoses were remeasured, according to the NASCET and ECST criteria. In total, of 147 cases, all above-mentioned parameters were obtained. Student's t tests for paired samples were used to evaluate the results. FINDINGS: The t-tests indicated significant differences between the per cent stenosis as measured on the anatomical specimen and on the angiogram (p<0.05). The results indicate that the angiographic measurement underestimates the degree of in-situ anatomical stenosis. The underestimation was more marked the less the degree of stenosis. CONCLUSIONS: Our study finds that per cent stenosis measurement obtained by angiography with NASCET or ECST methods does not reliably reflect the anatomical degree of per cent stenosis, which makes questionable the rigorous following of percentage stenosis using angiography as the sole indicator for carotid endarterectomy in all cases.

• MeSH
• algoritmy MeSH
• digitální subtrakční angiografie MeSH
• karotická endarterektomie MeSH
• lidé MeSH
• lineární modely MeSH
• reprodukovatelnost výsledků MeSH
• retrospektivní studie MeSH
• stenóza arteria carotis chirurgie   patologie   radiografie   MeSH
• stupeň závažnosti nemoci MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Remodeling of a microvascular network is common part of pathological changes associated with wide spectrum of diseases. Quantitative analysis of these alterations relies often on analysis of a point-pattern on the histological slide, i.e. on sections through the microvascular network only. Common techniques are based on the estimation of the average density of points representing section through microvessels on the histological image. This approach inherently omits the information about the regularity of the pattern. Thus, we used approach based on the Voronoi segmentation and chose the best statistical model of areas of Voronoi cells surrounding microvessels on 20 samples of human myocardium. The best model is based on the log-normal distribution. Parameters of the model for given data can be estimated as a mean and a standard deviation of logarithms of areas of Voronoi cells. Moreover, these parameters can be transformed to the widely used measure called the microvascular density.

BACKGROUND: Patients' life expectancy, clinical symptomatology and the extent of carotid stenosis are the most important factors when deciding whether to perform carotid endarterectomy (CEA) in patients with carotid stenosis. Therefore, the accuracy of measuring carotid stenosis is of utmost importance. METHODS: Patients with internal carotid artery (ICA) stenosis were investigated by digital subtraction angiography (DSA), Doppler ultrasonography (DUS) and magnetic resonance angiography (MRA). Atherosclerotic plaque specimens were transversally cut into smaller segments and histologically processed. The slides were scanned and specimens showing maximal stenosis were determined; the minimal diameter and the diameter of the whole plaque were measured. DSA, DUS and MRA measurements were obtained in 103 patients. A comparison between preoperative and histological findings was performed. In addition, correlation coefficients were computed and tested. RESULTS: Results show a significant correlation for each of the diagnostic procedures. Mean differences in the whole cohort between preoperative measurements and the histological measurements are as follows: angiographic measurement of carotid stenosis underestimated histological measurement by 14.5% and MRA by 0.7%, but DUS overestimated by 6.6%. The results in severe stenosis (> or =70%) are as follows: angiographic measurement underestimated the histological measurements by 2.3%, but MRA overestimated by 12.1% and DUS by 11.3%. The results in moderate stenosis (50-69%): angiographic measurement underestimated the histological measurements by 12.3%, but MRA overestimated by 0.2% and DUS by 7.2%. The results in mild stenosis (30-49%): angiographic measurement underestimated the histological measurements by 24.7% and MRA by 7.6%, but DUS overestimated by 3.3%. CONCLUSIONS: Our study confirms that DSA underestimates moderate and mild ICA stenosis. DUS slightly overestimated moderate ICA stenosis and highly overestimated high-grade ICA stenosis. MRA proved to be accurate in detecting moderate ICA stenosis, but slightly underestimated mild stenosis and overestimated high-grade stenosis. The surgeon should be aware of these discrepancies when deciding whether to perform CEA in patients with ICA stenosis.

• MeSH
• arteriae carotides * patologie   radiografie   ultrasonografie   MeSH
• digitální subtrakční angiografie metody   normy   MeSH
• kohortové studie MeSH
• lidé MeSH
• magnetická rezonanční angiografie metody   normy   MeSH
• prediktivní hodnota testů MeSH
• předoperační péče metody   MeSH
• stenóza arteria carotis diagnóza   patologie   ultrasonografie   MeSH
• ultrasonografie dopplerovská metody   normy   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• srovnávací studie MeSH