A comprehensive textbook of radiotherapy and related radiation physics and oncology for use by all those concerned with the uses of radiation and cytotoxic drugs in the treatment of patients with malignant disease. The first part covers underlying principles of physics, and the second is a systematic review by tumour site concentrating on the role of radiotherapy in the treatment of malignant disease and setting its use in context with chemotherapy and surgery.

• MeSH
• nádory radioterapie   MeSH
• nukleární lékařství metody   MeSH
• radioterapie metody   MeSH
• Publikační typ
• učebnice MeSH

• Konspekt
• Učební osnovy. Vyučovací předměty. Učebnice
• Lékařské vědy. Lékařství
• NLK Obory
• radiologie, nukleární medicína a zobrazovací metody
• onkologie
• NLK Publikační typ
• kolektivní monografie

OBJECTIVE: This study aimed to clarify the molecular epidemiology of hearing loss by identifying the responsible genes in patients without GJB2 mutations. STUDY DESIGN: Prospective genetic study. SETTING: Tertiary referral hospital. PATIENTS: Fifty one patients with bilateral sensorineural hearing loss, 20 men, and 31 women, mean age 24.9 years, range 3 to 64 years, from 49 families. GJB2 and deltaGJB6-D13S1830 mutations were excluded previously. INTERVENTION: Diagnostic. Sixty-nine genes reported to be causative of hearing loss were analyzed. Sequence capture technology, next-generation sequencing, and multiplex ligation-dependent probe amplification (MLPA) were used. Coverage of STRC was screened in Integrative Genomics Viewer software. MAIN OUTCOME MEASURE: Identification of causal pathogenic mutations in genes related to deafness. RESULTS: Five families (10%) had recessive STRC deletions or mutations. Five unrelated patients (10%) had recessive mutations in TMPRSS3, USH2A, PCDH15, LOXHD1, and MYO15A. Three families (6%) had autosomal dominant mutations in MYO6A, KCNQ4, and SIX1. One family (2%) had an X-linked POU3F4 mutation. Thus, we identified the cause of hearing loss in 28% of the families studied. CONCLUSIONS: Following GJB2, STRC was the second most frequently mutated gene in patients from the Czech Republic with hearing loss. To decrease the cost of testing, we recommend STRC deletion screening with MLPA before next-generation sequencing. The existence of a pseudogene and polymorphic STRC regions can lead to false-positive or false-negative results when copy number variation analysis is based on next-generation sequencing data.

• MeSH
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• membránové proteiny genetika   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mutace MeSH
• percepční nedoslýchavost vrozené   genetika   MeSH
• předškolní dítě MeSH
• prospektivní studie MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• předškolní dítě MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH

UNLABELLED: Pluripotent stem cell-derived committed neural precursors are an important source of cells to treat neurodegenerative diseases including spinal cord injury. There remains an urgency to identify markers for monitoring of neural progenitor specificity, estimation of neural fate and follow-up correlation with therapeutic effect in preclinical studies using animal disease models. Cell surface capture technology was used to uncover the cell surface exposed N-glycoproteome of neural precursor cells upon neuronal differentiation as well as post-mitotic mature hNT neurons. The data presented depict an extensive study of surfaceome during neuronal differentiation, confirming glycosylation at a particular predicted site of many of the identified proteins. Quantitative changes detected in cell surface protein levels reveal a set of proteins that highlight the complexity of the neuronal differentiation process. Several of these proteins including the cell adhesion molecules ICAM1, CHL1, and astrotactin1 as well as LAMP1 were validated by SRM. Combination of immunofluorescence staining of ICAM1 and flow cytometry indicated a possible direction for future scrutiny of such proteins as targets for enrichment of the neuronal subpopulation from mixed cultures after differentiation of neural precursor cells. These surface proteins hold an important key for development of safe strategies in cell-replacement therapies of neuronal disorders. BIOLOGICAL SIGNIFICANCE: Neural stem and/or precursor cells have a great potential for cell-replacement therapies of neuronal diseases. Availability of well characterised and expandable neural cell lineage specific populations is critical for addressing such a challenge. In our study we identified and relatively quantified several hundred surface N-glycoproteins in the course of neuronal differentiation. We further confirmed the abundant changes for several cell adhesion proteins by SRM and outlined a strategy for utilisation of such N-glycoproteins in antibody based cell sorting. The comprehensive dataset presented here demonstrates the molecular background of neuronal differentiation highly useful for development of new plasma membrane markers to identify and select neuronal subpopulation from mixed neural cell cultures.

• MeSH
• buněčná diferenciace fyziologie   MeSH
• buněčné linie MeSH
• kultivované buňky MeSH
• lidé MeSH
• membránové glykoproteiny metabolismus   MeSH
• nervové kmenové buňky cytologie   metabolismus   MeSH
• proteiny nervové tkáně metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Dokumentácia nálezov na prednom segmente oka a očnom pozadí pomocou prístrojov, ktoré umožňujú kvalitnú a precíznu diagnostiku, je dnes bežnou a dôležitou súčasťou skríningových projektov a je podstatnou súčasťou pri diagnostike, monitoringu a manažmente očných ochorení. V málo rozvinutých krajinách v rámci skríningových vyšetrení nemá oftalmológ k dispozícii moderné technológie ako biomikroskop alebo fundus kamera, ktoré by umožnili zobraziť predný alebo zadný segment oka na rovnakej úrovni ako je štandardom v rozvinutých krajinách. Cieľom našej práce je prezentovať prvé skúsenosti s fotodokumentáciou predného segmentu oka pomocou digitálneho fotoaparátu aj smartfónu a možnosti dokumentácie nálezov očného pozadia pomocou 20D Volkovej sférickej šošovky a smartfónu v rámci misijných projektov skríningu očných ochorení v Rwande a v Južnom Sudáne. Materiál a metodika: V rámci projektov skríningu očných ochorení v spolupráci s VŠZaSP sv. Alžbety v r. 2014 v Bigugu, Rwanda a v r. 2015 v Mapuordit, Južný Sudán, sme vyšetrovali pacientov v odľahlých častiach krajiny, ktorí nemali prístup k oftalmologickej starostlivosti. K dispozícii sme mali baterku, priamy oftalmoskop, tabuľky na zistenie zrakovej ostrosti pre analfabetov, Schiotzov tonometer, Volkovu šošovku, smartfón. Pacienti, ktorí absolvovali vyšetrenie, a potrebovali pomôcku – okuliare, dostali zo zbierky okuliarov už použité dioptrické okuliare alebo slnečné okuliare. Dokumentáciu nálezov na prednom segmente oka sme realizovali pomocou digitálneho fotoaparátu a u pacientov, u ktorých bolo potrebné dokumentovať nálezy očného pozadia zistené priamou oftalmoskopiou, sme využili možnosti smartfónu s fotoaparátom 8 Mpix a LED bleskom a Volkovej šošovky s hodnotou plus 20 dioptrií. Výsledky: V r. 2014 v Bigugu, Rwanda a v r. 2015 v Mapuorditte, Južný Sudán sme vyšetrovali pacientov v improvizovanej ambulancii bez dostupnosti elektrickej energie. Vyšetrili sme v r. 2014 celkove 340 pacientov a v r. 2015 celkove 290 pacientov. Vek pacientov bol vzhľadom na nedostupnosť identifikačných záznamov približne určný s pomocou tlmočníka, v oboch skupinách bol priemerný vek vyšetrených pacientov okolo 30 rokov. Najčastejšie ochorenia, ktoré viedli k praktickej slepote, boli katarakta, trachóm, poúrazové stavy. Infekčné ochorenia a následky neliečných infekčných ochorení boli v 20 % príčinou trvalých zmien na povrchu oka alebo pomocných orgánoch. V skupine pacientov HIV pozitívnych sme nezaznamenali patologické nálezy na očnom pozadí. Záver: Vyšetrenie predného segmentu oka štandardným digitálnym fotoaparátom a dokumentácia očného pozadia použitím smartfónu a Volkovej šošovky s hodnotou plus 20D je nenáročnou a zvládnuteľnou technikou, ktorou je možné zachytiť kvalitné a reprodukovateľné snímky vhodné na fotodokumentáciu a skríning v rámci projektov v ťažkých podmienkach v málo rozvinutých krajinách subsaharskej Afriky. V náročných klimatických aj geografických podmienkach v Rwande a v Južnom Sudáne sme vyšetrili pacientov, poskytli základnú pomoc odovzdaním použitej refrakčnej pomôcky zo charitatívnej zbierky, ktorú sme uskutočnili pred realizáciou projektu. Riešenie infekčných ochorení bolo možné len čiastočne (trachóm) vzhľadom na krátkosť trvania projektov. Možnosť využitia smartfónu s fotoaparátom 8 Mpix a LED bleskom a Volkovej šošovky s hodnotou plus 20 dioptrií je veľkým prínosom v takýchto projektoch pri vyšetrení očných ochorení v rozvojových krajinách.

Documentation of the anterior segment and the eye fundus with instruments that enable quality precision diagnostics is a common and important part of screening in humanitarian ophthalmology projects. It is the essential element in diagnosis, monitoring and management of eye diseases. In sub saharan countries within the screening for ophthalmologist are not available the modern technologies such as biomicroscope (slit lamp) or fundus camera. We describe our experience with photographs of anterior segment of the eye by using digital camera and Smartphone. The documentation of the eye fundus was recorded through 20D Volk spherical lens to Smartphone. Material and methods: Within the screening projects in collaboration with St. Elisabeth University of Health and Social Sciences for eye diseases in the year 2014 in Bigugu, Rwanda and in 2015 in Mapuordit, South Sudan, we examined patients who were unable to reach ophthalmologic care. We used a flashlight, a direct ophthalmoscope, tables to determine visual acuity on illiterate, Schi?tz tonometer, Volk lens, Smartphone. Patients who underwent screening, and needed glasses, got from humanitarian collection already used dioptric eyeglasses or sunglasses. For documentation of the anterior segment we used a digital camera and for patients in whom it was necessary to document fundus findings detected by direct ophthalmoscopy we took the opportunity of Smartphone with 8 Mpix camera and the LED flash and Volk lens plus 20 Diopters. Results: In 2014 within the project in Bigugu, Rwanda and in 2015 in Mapuordit, South Sudan, we examined patients in an improvised clinic without access to electricity. We examined in 2014 a total of 340 patients and in 2015 a total of 290 patients. Patient age was due to the unavailability of designated identification records estimated with the help of an interpreter. In both groups, the mean age of the patients was about 30 years. The most common diseases leading to blindness were cataract, trachoma, post-traumatic conditions. Infectious diseases and consequences of untreated infectious diseases were the cause of 20% of the permanent changes on the surface of the eye or the adnexa. In the group of HIV positive patients we did not mention pathological findings on the eye fundus. Conclusion: Anterior segment findings documentation with digital camera or mobile phone and fundus examination using a Smartphone and Volks lens with a value of plus 20D is inexpensive and manageable technique which can capture high quality and reproducible images. These techniques are suitable for photo documentation of anterior segment and also eye fundus screening within humanitarian projects of eye diseases in developing countries.

• MeSH
• chytrý telefon * využití   MeSH
• design vybavení MeSH
• diagnostické techniky oftalmologické přístrojové vybavení   MeSH
• dobrovolné zdravotnické agentury MeSH
• dokumentace metody   MeSH
• dospělí MeSH
• dostupnost zdravotnických služeb MeSH
• fotografování * metody   přístrojové vybavení   MeSH
• lidé MeSH
• mobilní telefon využití   MeSH
• oční nemoci * diagnóza   prevence a kontrola   MeSH
• oftalmoskopie metody   MeSH
• rozvojové země MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• Geografické názvy
• Jižní Súdán MeSH
• Rwanda MeSH

Cell surface proteins are major targets of biomedical research due to their utility as cellular markers and their extracellular accessibility for pharmacological intervention. However, information about the cell surface protein repertoire (the surfaceome) of individual cells is only sparsely available. Here, we applied the Cell Surface Capture (CSC) technology to 41 human and 31 mouse cell types to generate a mass-spectrometry derived Cell Surface Protein Atlas (CSPA) providing cellular surfaceome snapshots at high resolution. The CSPA is presented in form of an easy-to-navigate interactive database, a downloadable data matrix and with tools for targeted surfaceome rediscovery (http://wlab.ethz.ch/cspa). The cellular surfaceome snapshots of different cell types, including cancer cells, resulted in a combined dataset of 1492 human and 1296 mouse cell surface glycoproteins, providing experimental evidence for their cell surface expression on different cell types, including 136 G-protein coupled receptors and 75 membrane receptor tyrosine-protein kinases. Integrated analysis of the CSPA reveals that the concerted biological function of individual cell types is mainly guided by quantitative rather than qualitative surfaceome differences. The CSPA will be useful for the evaluation of drug targets, for the improved classification of cell types and for a better understanding of the surfaceome and its concerted biological functions in complex signaling microenvironments.

• MeSH
• buněčné linie MeSH
• databáze proteinů MeSH
• hmotnostní spektrometrie metody   MeSH
• lidé MeSH
• membránové proteiny chemie   MeSH
• myši MeSH
• proteomika metody   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

The most promising near-term application of circulating tumor cells (CTCs) monitoring relates to the development of targeted cancer therapies, and the need to tailor such treatments to individual tumor characteristics. A high number of new innovative technologies to improve methods for detecting CTCs, with extraordinarily high sensitivity, have recently been presented. The identification and characterization of CTCs require extremely sensitive and specific methods that are able to isolate CTCs with the possibility of cultivation and downstream analysis of in vitro culture of separated CTCs. In this original research paper, we demonstrate that it is possible to isolate human CTCs from a patient with prostate cancer, with subsequent cultivation and proliferation in vitro. We show that the use of a filtration device implemented by MetaCell® can fulfil all the requirements mentioned above. Fifty-five patients with localized prostate cancer have so far been enrolled into the study. CTCs were detected in the blood samples of 28 (52%) out of the 55 patients. We report successful isolation of CTCs from patients with prostate cancer, capturing cells with a proliferative capacity in 18 (64.3%) out of the 28 CTC-positive patients. Direct correlation with Gleason score and T stage was not proven. The cells, captured by a size-based filtration approach, remain in a good state, unaffected by any antibodies or lysing solutions. During the filtration process, no interactions occurred between antibodies and antigens on the surface of CTCs. This biological interaction is specific for immunomagnetic methods. The MetaCell device provides the possibility of reaching virgin CTCs suitable for subsequent cultivation or single-cell analysis. This aspect will have an important impact on the future design of clinical trials testing new drugs against targets expressed on metastatic cancer cells. In addition to measurement of CTC counts, future trials with targeted therapies should also include the assessment of the specific therapeutic target on CTCs.

• MeSH
• cytologické techniky metody   MeSH
• kohortové studie MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádorové buňky kultivované MeSH
• nádorové cirkulující buňky patologie   MeSH
• nádory prostaty krev   patologie   MeSH
• senioři MeSH
• staging nádorů MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The combination of microarray technologies with microfluidic sample delivery and real-time detection methods has the capability to simultaneously monitor 10-1000 s of biomolecular interactions in a single experiment. Despite the benefits that microfluidic systems provide, they typically operate in the laminar flow regime under mass transfer limitations, where large analyte depletion layers act as a resistance to analyte capture. By locally stirring the fluid and delivering fresh analyte to the capture spot, the use of passive mixing structures in a microarray environment can reduce the negative effects of these depletion layers and enhance the sensor performance. Despite their large potential, little attention has been given to the integration of these mixing structures in microarray sensing environments. In this study, we use passive mixing structures to enhance the mass transfer of analyte to a capture spot within a microfluidic flow cell. Using numerical methods, different structure shapes and heights were evaluated as means to increase local fluid velocities, and in turn, rates of mass transfer to a capture spot. These results were verified experimentally via the real-time detection of 20-mer ssDNA for an array of microspots. Both numerical and experimental results showed that a passive mixing structure situated directly over the capture spot can significantly enhance the binding rate of analyte to the sensing surface. Moreover, we show that these structures can be used to enhance mass transfer in experiments regarding an array of capture spots. The results of this study can be applied to any experimental system using microfluidic sample delivery methods for microarray detection techniques.

• MeSH
• biosenzitivní techniky přístrojové vybavení   MeSH
• design vybavení MeSH
• difuze MeSH
• jednovláknová DNA analýza   MeSH
• mikročipová analýza přístrojové vybavení   MeSH
• mikrofluidní analytické techniky přístrojové vybavení   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

A better description of the leukemia cell surface proteome (surfaceome) is a prerequisite for the development of diagnostic and therapeutic tools. Insights into the complexity of the surfaceome have been limited by the lack of suitable methodologies. We combined a leukemia xenograft model with the discovery-driven chemoproteomic Cell Surface Capture technology to explore the B-cell precursor acute lymphoblastic leukemia (BCP-ALL) surfaceome; 713 cell surface proteins, including 181 CD proteins, were detected through combined analysis of 19 BCP-ALL cases. Diagnostic immunophenotypes were recapitulated in each case, and subtype specific markers were detected. To identify new leukemia-associated markers, we filtered the surfaceome data set against gene expression information from sorted, normal hematopoietic cells. Nine candidate markers (CD18, CD63, CD31, CD97, CD102, CD157, CD217, CD305, and CD317) were validated by flow cytometry in patient samples at diagnosis and during chemotherapy. CD97, CD157, CD63, and CD305 accounted for the most informative differences between normal and malignant cells. The ALL surfaceome constitutes a valuable resource to assist the functional exploration of surface markers in normal and malignant lymphopoiesis. This unbiased approach will also contribute to the development of strategies that rely on complex information for multidimensional flow cytometry data analysis to improve its diagnostic applications.

• MeSH
• akutní lymfatická leukemie imunologie   metabolismus   MeSH
• CD antigeny analýza   MeSH
• imunofenotypizace MeSH
• lidé MeSH
• membránové proteiny * analýza   metabolismus   MeSH
• myši MeSH
• nádorové biomarkery * analýza   MeSH
• proteom * analýza   metabolismus   MeSH
• průtoková cytometrie MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH