Úvod: Histologické vyšetrenie počas operácie (RR) má svoje miesto v chirurgickom manažmente diferencovaného karcinómu štítnej žľazy (DTC). Rozšírenie indikácie k limitovanému zákroku na 4cm veľkosti tumoru (ATA guidelines 2015) cytologicky verifikovaných DTC, zvyšuje dôraz na akurátnu selekciu pacientov. V našej práci sme sa zamysleli nad efektivitou RR a jeho vzťahu k optimálnemu manažmentu pacientov. Metódy: V retrospektívnej štúdii sme vyhodnotili dokumentáciu pacientov v OÚSA indikovaných na primárnu operáciu pre DTC od 1. 1. 2016 do 31. 12. 2020 – 489 pacientov, 121 mužov, medián veku 50 rokov (16−81), 73 pacientov (ženské pohlavie, vek 18−45 rokov) s predoperačne identifikovaným nízkorizikovým DTC (veľkosť 11−40mm) bolo indikovaných k lobektómii. Výsledky: Kritéria pre limitovaný zákrok nesplnilo 34 pacientov (46,6 %) − 15 pacientov bolo identifikovaných z RR z lymfatickej uzliny centrálneho kompartmentu (LUCK) (15 z 25 pacientov) − falošne negatívny nález 1 pacient, 6 pacienti RR zo štítnej žľazy (ŠŽ) (6/41) – falošne negatívny nález 11 pacienti. Dvojkroková operácia OP bola vykonaná u 13 pacientov (17,8 %). RR z LUCK štatisticky významne identifikoval rizikový karcinóm a znížil riziko dvojkrokovej operácie (OR 1,93, p= 0,026), oproti skupine pacientov, u ktorých RR nebol alebo bol vykonaný zo ŠŽ. Záver: Približne len polovica pacientov z predoperačne identifikovaných nízkorizikových karcinómov v našom súbore splnilo kritéria pre limitovaný zákrok. Z nich cca 30 % malo dvojkrokovú operáciu. Peroperačné vyšetrenie LUCK umožnilo vykonať radikálny zákrok v jednej dobe.
Introduction: Histological examination during surgery (FS) has a place in the surgical management of differentiated thyroid carcinoma (DTC). Extending the indication for limited surgery to 4 cm tumor size (ATA guidelines 2015) cytologically verified DTCs, increases the emphasis on accurate patient selection. In our work, we reflected on the effectiveness of FS and its relationship to optimal patient management. Methods: In a single-center retrospective study, we evaluated the documentation of patients indicated for primary surgery for DTC from January 1, 2016 to December 31, 2020 – there was 489 patients collectively, 121 were men, median age was 50 years (16−81), 73 patients (female, age 18−45 years) with preoperatively identified low-risk DTC (size 11−40mm) were indicated for lobectomy. Results: 34 patients (46.6%) did not meet the criteria for limited surgery − 15 patients were identified from FS of the lymph nodes of the central compartment (LNCK) (15 of 25 patients) – 1 patient with false negative result and 6 patients with FS of the thyroid gland (SH) (6 / 41) – 11 patients with false negative findings. Two-step OP surgery was performed on 13 patients (17.8%). FS of LNCK identified high-risk cancer and reduced the risk of two-step surgery compared to the group of patients in whom FS was not performed or was performed from thyroid gland. The difference was statistically significant (OR 1.93, p=0.026). Conclusion: Approximately ½ of the patients from preoperatively identified low-risk cancers in our cohort met the criteria for limited surgery. About 30% of them eventually needed a two-step operation. Perioperative examination of LNCK helps to perform radical surgery at one time.
- MeSH
- adenokarcinom patologie MeSH
- histologické techniky MeSH
- krk patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- lymfatické metastázy MeSH
- lymfatické uzliny * chirurgie patologie MeSH
- nádory štítné žlázy * chirurgie patologie MeSH
- peroperační doba MeSH
- retrospektivní studie MeSH
- zmrazené řezy MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
CONTEXT: The question of the ability of frozen section analysis (FSA) to accurately detect malignant pathology intraoperatively has been discussed for many decades. OBJECTIVE: We aimed to conduct a systematic review and meta-analysis assessing the diagnostic estimates of FSA of the urethral and ureteral margins in patients treated with radical cystectomy (RC) for bladder cancer (BCa). EVIDENCE ACQUISITION: The MEDLINE and EMBASE databases were searched in February 2021 for studies analyzing the association between FSA and the final urethral and ureteral margin status in patients treated with RC for BCa. The primary endpoint was the value of pathologic detection of urethral and ureteral malignant involvement with FSA during RC compared with the final margin status. We included studies that provided true positive, true negative, false positive, and false negative values for FSA, which allowed us to calculate the diagnostic estimates. EVIDENCE SYNTHESIS: Fourteen studies, comprising 8208 patients, were included in the quantitative synthesis. Forest plots revealed that the pooled sensitivity and specificity for FSA of urethral margins during RC were 0.83 (95% confidence interval [CI] 0.38-0.97) and 0.95 (95% CI 0.91-0.97), respectively. While for the FSA of ureteral margins, the pooled sensitivity and specificity were 0.77 (95% CI 0.67-0.84) and 0.97 (95% CI 0.95-0.98), respectively. Calculated diagnostic odds ratios indicated high FSA effectiveness, and patients with a positive urethral or ureteral margin at final pathology are over 100 times more likely to have positive FSA than patients without margin involvement at final pathology. Area under the curves of 96.6% and 96.7% were reached for FSA detection of urethral and ureteral tumor involvement, respectively. CONCLUSIONS: Intraoperative FSA demonstrated high diagnostic performance in detecting both urethral and ureteral malignant involvement at the time of RC for BCa. FSA of both urethral and ureteral margins during RC is accurate enough to be of great value in the routine management of BCa patients treated with RC. While its specificity was great to guide intraoperative decision-making, its sensitivity remains suboptimal yet. PATIENT SUMMARY: We believe that the frozen section analysis of both urethral and ureteral margins during radical cystectomy should be considered more often in urologic practice, until quality of life-based cost-effectiveness studies can identify patients within each institution who are unlikely to benefit from it.
- MeSH
- cystektomie MeSH
- kvalita života MeSH
- lidé MeSH
- nádory močového měchýře * diagnóza patologie chirurgie MeSH
- resekční okraje MeSH
- ureter * patologie chirurgie MeSH
- zmrazené řezy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- metaanalýza MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- systematický přehled MeSH
The disconnect between preclinical and clinical results underscores the imperative for establishing good animal models, then gleaning all available data on efficacy, safety, and potential toxicities associated with a device or drug. Mini pigs are a commonly used animal model for testing orthopedic and dental devices because their skeletons are large enough to accommodate human-sized implants. The challenge comes with the analyses of their hard tissues: current methods are time-consuming, destructive, and largely limited to histological observations made from the analysis of very few tissue sections. We developed and employed cryo-based methods that preserved the microarchitecture and the cellular/molecular integrity of mini pig hard tissues, then demonstrated that the results of these histological, histochemical, immunohistochemical, and dynamic histomorphometric analyses e.g., mineral apposition rates were comparable with similar data from preclinical rodent models. Thus, the ability to assess static and dynamic bone states increases the translational value of mini pig and other large animal model studies. In sum, this method represents logical means to minimize the number of animals in a study while simultaneously maximizing the amount of information collected from each specimen.
- MeSH
- fyziologická kalcifikace MeSH
- kryoprezervace metody MeSH
- kryoultramikrotomie metody MeSH
- lebka cytologie MeSH
- miniaturní prasata MeSH
- odběr biologického vzorku metody MeSH
- polyethylenglykoly MeSH
- prasata MeSH
- remodelace kosti MeSH
- sacharosa MeSH
- sodná sůl karboxymethylcelulosy MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
The gold standard for the treatment of periocular basal cell carcinoma is surgical resection followed by ophthalmoplastic reconstruction. The highest priority in most cases is the complete histopathologically controlled tumor excision. The histopathological preparation can be carried out in two stages by rapid overnight embedding or intraoperatively by a rapid frozen section procedure. A variety of reconstruction methods enable a customized and in most cases also a cosmetically and functionally attractive defect coverage. Postoperatively, a regularly performed tumor aftercare is essential.
- MeSH
- bazocelulární karcinom * MeSH
- lidé MeSH
- nádory kůže * MeSH
- nádory očního víčka MeSH
- retrospektivní studie MeSH
- zmrazené řezy MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
Siderophores represent important microbial virulence factors and infection biomarkers. Their monitoring in fermentation broths, bodily fluids, and tissues should be reproducible. Similar isolation, characterization, and quantitation studies can often have conflicting results, and without proper documentation of sample collection, data processing, and analysis methods, it is difficult to reexamine the data and reconcile these differences. In this Springer Nature Protocol, we present the procedure optimized for ferricrocin/triacetylfusarinine C extraction from biological material as well as for tissue fixation and cryosectioning for optical microscopy and for both elemental and molecular mass spectrometry imaging. Special attention is paid to siderophore data mining from conventional and product ion mass spectra, liquid chromatography, and mass spectrometry imaging datasets, performed here by our free software called CycloBranch.
- MeSH
- Aspergillus fumigatus metabolismus MeSH
- biologické markery analýza MeSH
- chromatografie kapalinová metody MeSH
- data mining metody MeSH
- datové soubory jako téma MeSH
- ferrichrom analogy a deriváty izolace a purifikace metabolismus MeSH
- fixace tkání metody MeSH
- hmotnostní spektrometrie metody MeSH
- invazivní plicní aspergilóza diagnóza mikrobiologie MeSH
- kryoultramikrotomie metody MeSH
- krysa rodu rattus MeSH
- kyseliny hydroxamové izolace a purifikace metabolismus MeSH
- lidé MeSH
- modely nemocí na zvířatech MeSH
- siderofory izolace a purifikace metabolismus MeSH
- software MeSH
- železité sloučeniny izolace a purifikace metabolismus MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
There are various preparatory techniques for light microscopy permitting access to the inner structure of plant body and its development. Minute objects might be processed as whole-mount preparations, while voluminous ones should be separated into smaller pieces. Here we summarize some of the "classical" techniques to cut more voluminous objects into slices and access their inner structure either for simple anatomical analysis or for further processing (e.g., histochemistry, immunohistochemistry, in situ hybridization, enzyme histochemistry).
- MeSH
- barvení a značení metody MeSH
- fixace tkání metody MeSH
- histocytochemie metody MeSH
- histocytologické preparační techniky metody MeSH
- mikroskopie metody MeSH
- mikrotomie metody MeSH
- rostliny chemie ultrastruktura MeSH
- zalévání tkání do parafínu metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The findings obtained by the famous nineteenth-century Czech scientist Jan Evangelista Purkyně (1787-1869) in the field of microscopic structure of animal and human tissues, including the brain, spinal cord, and nerves, have already been described in depth in a number of older and newer publications. The present article contains an overview of the instruments and tools that Purkyně and his assistants used for microscopic research of tissue histology. Some of these instruments were developed either by Purkyně alone, such as the microtomic compressor, or together with his assistant Adolph Oschatz, such as the microtome. A brief overview of the development of the cutting engines suggests that the first microtome, a prototype of modern sliding microtomes, was designed and constructed under the supervision of Purkyně at the Institute of Physiology in Wrocław. Purkyně and his assistants, thus, not only obtained important findings of animal and human nervous and other tissues but also substantially contributed to the development of instruments and tools for their study, a fact often forgotten today.
- MeSH
- dějiny 19. století MeSH
- lidé MeSH
- mikroskopie dějiny přístrojové vybavení MeSH
- mikrotomie dějiny přístrojové vybavení MeSH
- neurovědy dějiny přístrojové vybavení MeSH
- výzkum dějiny přístrojové vybavení MeSH
- zvířata MeSH
- Check Tag
- dějiny 19. století MeSH
- lidé MeSH
- zvířata MeSH
- Publikační typ
- biografie MeSH
- časopisecké články MeSH
- historické články MeSH
- Geografické názvy
- Československo MeSH
- O autorovi
- Purkyně, Jan Evangelista, 1787-1869 Autorita
We present a new method of multiple immunolabeling that is suitable for a broad spectrum of biomedical applications. The general concept is to label both sides of the ultrathin section with the thickness of 70-80 nm with different antibodies conjugated to gold nanoparticles and to distinguish the labeled side by advanced imaging methods with high resolution scanning electron microscopy, such as by correlating images acquired at different energies of primary electrons using different signals. From the Clinical Editor: The use of transmission electron microscopy has become an indispensible tool in the detection of cellular proteins. In this short but interesting article, the authors described their new method of labeling and the identification of four different proteins simultaneously, which represents another advance in imaging technique.
- MeSH
- akrylové pryskyřice chemie MeSH
- barvení a značení metody MeSH
- imunohistochemie MeSH
- kovové nanočástice chemie ultrastruktura MeSH
- mikrotomie metody MeSH
- reprodukovatelnost výsledků MeSH
- senzitivita a specificita MeSH
- skenovací elektrochemická mikroskopie metody MeSH
- vylepšení obrazu metody MeSH
- zlato chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Scanning electron microscopes are useful biological tools that can be used to image the surface of whole organisms, tissues, cells, cellular components, and macromolecules. Processes and structures that exist at surfaces can be imaged in pseudo, or real 3D at magnifications ranging from about 10× to 1,000,000×. Therefore a whole multicellular organism, such as a fly, or a single protein embedded in one of its cell membranes can be visualized. In order to identify that protein at high resolution, or to see and quantify its distribution at lower magnifications, samples can be labeled with antibodies. Any surface that can be exposed can potentially be studied in this way. Presented here is a generic method for immunogold labeling for scanning electron microscopy, using two examples of specimens: isolated nuclear envelopes and the cytoskeleton of mammalian culture cells. Various parameters for sample preparation, fixation, immunogold labeling, drying, metal coating, and imaging are discussed so that the best immunogold scanning electron microscopy results can be obtained from different types of specimens.
- MeSH
- antigeny genetika metabolismus MeSH
- barvení a značení metody MeSH
- buněčná membrána metabolismus ultrastruktura MeSH
- cytoskelet metabolismus ultrastruktura MeSH
- epoxidové pryskyřice chemie MeSH
- exprese genu MeSH
- fixace tkání metody MeSH
- fixativa chemie MeSH
- formaldehyd chemie MeSH
- imunohistochemie metody MeSH
- jaderný obal metabolismus ultrastruktura MeSH
- koloidní zlato chemie MeSH
- komplex proteinů jaderného póru genetika metabolismus MeSH
- mikroskopie elektronová rastrovací metody MeSH
- mikrotomie MeSH
- oocyty metabolismus ultrastruktura MeSH
- polymery chemie MeSH
- protilátky chemie MeSH
- Xenopus laevis MeSH
- zalévání tkání metody MeSH
- zvířata MeSH
- Check Tag
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Immunolabeling electron microscopy is a challenging technique with demands for perfect ultrastructural and antigen preservation. High-pressure freezing offers an excellent way to fix cellular structure. However, its use for immunolabeling has remained limited because of the low frequency of labeling due to loss of protein antigenicity or accessibility. Here we present a protocol for immunogold labeling of the yeast Saccharomyces cerevisiae that gives specific and multiple labeling while keeping the finest structural details. We use the protocol to reveal the organization of individual nuclear pore complex proteins and the position of transport factors in the yeast Saccharomyces cerevisiae in relation to actual transport events.
- MeSH
- barvení a značení metody MeSH
- epoxidové pryskyřice chemie MeSH
- exprese genu MeSH
- fixace tkání metody MeSH
- fixativa chemie MeSH
- glutaraldehyd chemie MeSH
- imunoelektronová mikroskopie metody MeSH
- imunohistochemie metody MeSH
- komplex proteinů jaderného póru genetika metabolismus MeSH
- kryoprezervace metody MeSH
- mikrotomie MeSH
- mrazová substituce metody MeSH
- protilátky chemie MeSH
- Saccharomyces cerevisiae - proteiny genetika metabolismus MeSH
- Saccharomyces cerevisiae metabolismus ultrastruktura MeSH
- zalévání tkání metody MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH