Úvod: Skupina neuroendokrinných tumorov vychádzajúca z buniek pankreasu sa nazýva pankreatické neuroendokrinné tumory (PNET). Kombinácia EUS a elastografie (EG) rozširuje diagnostické a zobrazovacie možnosti. Ciele: Cieľom našej práce bolo zistiť zastúpenie obrazov B-módu pre PNET, vyhodnotenie typického EG obrazu PNET, využitie „strain ratio“ (SR) a „strain histogram“ (SH) v diferenciálnej diagnostike PNET, stanovenie „cut off“ hodnoty SR a SH pre PNET a porovnanie štandardizovaných meraní s literatúrou. Metódy: Do štúdie boli zaradení pacienti vyšetrovaní na Internej klinike gastroenterologickej. Celkovo bolo vyšetrených 31 pacientov. Do súboru bolo zaradených 25 pacientov (8 mužov, 17 žien). Priemerný vek bol 52,76 roka (14–74). Všetkým zaradeným pacientom bolo realizované neinvazívne vyšetrenie endoskopická ultrasonografia. Po lokalizácii ložiska ultrasonograficky bol prvý záznam realizovaný po zmrazení obrazu v B-móde s vykonaním merania jeho veľkosti. Následne bolo realizované „strain“ elastografické meranie. V sledovanej skupine sme zaznamenali priemernú veľkosť 12,75 mm. Výsledky: Charakteristiky obrazu v B-móde pre PNET boli v 68 % hypoechogénne, v 12 % hyperechogénne, v 12 % izoechogénne a v 8 % zmiešanej echogenity. PNET boli v B-móde v 80 % ostro ohraničené a v 20 % neostro ohraničené. Presnosť priradenia elastografickej hodnoty typickej pre malígne nádory pankreasu bola pre PNET 96 % s použitím 5-stupňovej klasifikácie a 88 % s použitím 4-stupňovej klasifikácie. Senzitivita zaradenia PNET medzi malígne nádory bola pri „cut off“ SR > 3,2 80 % a SH < 50 100 %. Diskusia: Vzhľadom na priemerné veľkosti nádorov pozorované v našej štúdii, EUS prináša vysokú senzitivitu v diagnostike PNET v čase, kedy je u nich ešte možné aj miniinvazívne odstránenie. Okrem typického obrazu hypoechogenity a menej často popisovaných obrazov hyperechogenity a izoechogenity pre PNET, sme pozorovali aj echogenitu zmiešanú. Elastografický kvalitatívny obraz „strain“ hodnotený štvor- a päť- stupňovou klasifikáciou sa ukázal ako spoľahlivý pri odlíšení PNET a GNET od benígnych nádorov. V kvantitatívnej elastografii sú hodnoty SR a SH na pomedzí malígnych a benígnych ložísk pankreasu, jedným z dôvodou takýchto výsledkov môže byť rôznorodosť tejto skupiny nádorov pri rôznej mitotickej aktivite – „grade“. Záver: Konzistentnosť nami publikovaných výsledkov ukazuje použiteľnosť tejto metódy v prípade rozhodovania o definitívnej diagnóze, ak histológia nádoru nie je a nemôže byť dostupná.

Introduction: The group of neuroendocrine tumors derived from pancreatic cells is called pancreatic neuroendocrine tumors (PNETs). The combination of EUS and elastography (EG) expands diagnostic and imaging capabilities. Aim: The aim of our work was to determine the representative images of B-mode for PNET, evaluation of a typical EG image of PNETs, use of strain ratio (SR) and “strain histogram” (SH) in differential diagnosis of PNETs, determination of SR and SH cut off value for PNETs and comparison of standardized measurements with literature. Methods: Patients examined at the Internal Gastroenterology Clinic were included in the cohort. A total of 31 patients were examined. The group included 25 patients (8 men, 17 women). The mean age in group was 52.76 years (14–74). Non-invasive examination by endoscopic ultrasonography was performed on all patients. After locating the lesion by ultrasound, the first recording was made after freezing the image in B-mode and performing size measurement. Subsequently, a Strain elastography measurement was performed. In the monitored group we recorded an average size of 12.75 mm. Results: The characteristics of the image in B-mode were as follows for PNETs 68% hypoechogenic, 12% hyperechogenic, 12% isoechogenic and 8% mixed echogenicity. 80% of PNETs in B-mode were sharply demarcated and 20% with blurred borders. The accuracy of the value assignment typical of pancreatic malignancies using elastography was 96% for PNET in 5-degree classification system and 88% for a 4-degree classification, for SR cut off >3.2 with a sensitivity of 80% and SH cut off <50 100%. Discussion: Given the average tumor sizes observed in our study, EUS provides high sensitivity in PNET diagnostics and allows diagnostics at a time when minimally invasive removal is still possible. In addition to the typical picture of hypoechogenicity and the less frequently described pictures of hyperechogenicity and isoechogenicity for PNET, we also observed mixed echogenicity. The elastographic qualitative strain image evaluated by the four- and five- -degree classification proved to be reliable in distinguishing PNET from benign tumors. In quantitative elastography, the values of SR are between malignant and benign deposits of the pancreas, one of the reasons for such values may be the diversity of this group of diseases with different mitotic activity – grade of the tumor. Conclusion: Consistency of the results published by us, shows the applicability of this method in deciding on a definitive diagnosis if tumor histology is not and cannot be available.

• MeSH
• dospělí MeSH
• elastografie * MeSH
• endosonografie MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• nádory slinivky břišní diagnostické zobrazování   MeSH
• neuroendokrinní nádory * diagnostické zobrazování   MeSH
• pankreas anatomie a histologie   diagnostické zobrazování   MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH

Objev indukované pluripotence v roce 2006 umožnil revoluční způsob získávaní autologních terapeuticky aplikovatelných buněk, a mož‐ nost modelovat jakékoliv onemocnění v in vitro podmínkách. Možnost vrátit libovolnou, finálně diferencovanou buňku „v čase“ zpátky do stádia pluripotence je zajímavé i pro oblast onkologického výzkumu. Tato technologie umožnila studium procesů spojených s roz‐ vojem nádorového fenotypu buňky a taky s přechodem nádorové buňky do stádia s nižší mírou diferenciace. Reprogramování buněk do indukovaných pluripotentních kmenových buněk také pomáhá mnohem lépe studovat raritní populaci buněk, přítomných v nádo‐ rech – tzv. nádorové kmenové buňky. Indukovaná pluripotence některých typů nádorových buněk, spojená s jejich následnou řízenou diferenciací by se zároveň mohla stát jednou z možných terapeutických aplikací v onkologii.

Discovery of technology of induced pluripotency that allows the generation of autologous therapeutically applicable cells and generati‐ on of in vitro cell models for diseases with limited (or highly invasive) access to tested cells has also opened new horizons in the field of oncology research. The unique ability to reprogram the cancer cell into pluripotency with subsequent directed differentiation into cell with no malignant phenotype should be considered as a challenge in the field of new oncotherapy development. Although still conside‐ red to be realistic only on the level of experimental approach, the recent progress in the field of induced pluripotency gives the hope that dedifferentiation‐based therapies connected with the erase of malignant phenotype of original cancer cell will be more realistic in near future. By then, the most important role of induced pluripotency in oncology remains in the field of regenerative therapy as a source of autologous cells for regeneration of tissues or organs damaged by tumor growth or aggressive therapy

• MeSH
• lidé MeSH
• nádorové kmenové buňky MeSH
• nádory terapie   MeSH
• přeprogramování buněk MeSH
• techniky buněčného přeprogramování * metody   MeSH
• Check Tag
• lidé MeSH

Appendiceal mucocele is a rare disease with an incidence of 0.07-0.63% of all appendectomies and was first described in 1842 by Carl von Rokitansky. It is defined as an abnormal intraluminal accumulation of mucin. The clinical picture of AM can vary from asymptomatic mass in the right lower quadrant to symptoms of acute appendicitis. In some cases, AM can be found accidentally on CT performed due to other reasons or during surgery. Diagnosis consists mainly of imaging methods such as ultrasound, CT, and MRI with the finding of encapsulated cystic mass with calcifications. The main goal of surgical treatment is to remove an intact mucocele and prevent spillage of mucin into the peritoneal cavity. We present a case of large mucocele treated with laparoscopic right hemicolectomy.

• MeSH
• apendix * diagnostické zobrazování   chirurgie   MeSH
• kolektomie MeSH
• laparoskopie * MeSH
• lidé MeSH
• muciny MeSH
• mukokéla * diagnostické zobrazování   chirurgie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Pankreatický duktální adenokarcinom (PDAC) představuje jeden z nejagresivnějších typů lidských malignit. V současnosti je toto zhoubné onemocnění čtvrtou nejčastější příčinou úmrtí na rakovinu. Pětileté přežití pacientů s duktálním adenokarcinomem je méně než 8 %. Nové in vitro a in vivo modely jsou proto nutně potřebné pro vývoj nových terapií. S vlastní technologií pro derivaci nových, unikátních 3D nádorových buněčných linií izolovaných z lidských nádorů a ve spolupráci s Ústavem živočišné fyziologie a genetiky AV ČR v Liběchově představujeme plán vývoje velkého zvířecího modelu pro modelování lidského PDAC. S využitím tohoto modelu a nejmodernějších laboratorních technik provedeme profilovací analýzy (detekce a fenotypování cirkulujících rakovinných buněk, izolace a sekvenování cirkulující DNA, metabolomické profilování a analýzu onkoproteinů, a detekce cytokinů pomocí multiplexních protilátkových čipů za účelem nalezení biomarkerů nádoru pankreatu. Nově vyvinutý model zároveň poskytneme pro potřeby výzkumu spolupracujícím vědeckým pracovištím a farmaceutickým společnostem.

Pancreatic ductal adenocarcinoma represents one of the most aggressive type of human malignancy. Currently, this malignancy is the fourth most frequent cause of dead. 5-year survival of patients with ductal adenocarcinoma is less than 8 %. New in vitro and in vivo models are therefore desperately needed for new therapy development. With our own technology for derivation of new, unique 3D cancer cell lines from human tumors and in collaboration with Institute of Animal Physiology and Genetics, AS CR, in Libechov, we present here the plan for the development of large xenograft animal model (pig) bearing human pancreatic tumor. With the use of this model and state -of -the -art lab.techniques, we will perform multiple profiling analyses (circulating cancer cell detection and phenotyping, circulating DNA isolation and sequencing, metabolomic profiling and cancer -related proteins and cytokines detection with multiplex antibody array chips in order to find the hallmarks of pancreatic tumor.

One of the most common mechanisms of immune evasion in MSI colorectal cancers (CRCs) is loss of HLA class I expression due to mutations in B2M gene which can become a negative predictor for checkpoint blockade therapy. The aim of this study was the determination of prevalence of B2M somatic mutations in MSI CRC patients and relationship between B2M mutations and lymphocytes infiltration and other clinicopathological features as well as detection of methylation changes in B2M promoter region which can be another mechanism of immune escape. In our study, 37 MSI-H and 5 MSI-L patients were selected for screening of B2M mutational and methylation status. The characterization of patients was based on standard histopathological diagnosis and TNM classification; BRAF, KRAS mutations, tumor-infiltrating lymphocytes and peritumoral lymphoid reaction were also determined. MSI analysis was performed using fragment analysis. B2M mutations were identified by Sanger sequencing, and methylation of CpG islands in promoter region was detected by methylation-specific PCR. Heterozygous mutations in the B2M gene were detected in five MSI-H patients (13.5%), while the mutation c.45_48delTTCT was determined in four patients and mutation c.276delC was found in two patients. One of these five patients was compound heterozygote harboring both mutations. Methylation of the promoter region of the B2M gene was observed in one patient with MSI-H colorectal cancer. Detection of genetic and epigenetic changes in B2M gene could be important in personalized therapy for CRC patients as these changes may be one of the mechanisms of secondary resistance of MSI positive tumors to immunotherapy.

• MeSH
• beta-2-mikroglobulin genetika   MeSH
• CpG ostrůvky MeSH
• dospělí MeSH
• down regulace MeSH
• epigeneze genetická MeSH
• kolorektální nádory genetika   patologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• metylace DNA * MeSH
• mikrosatelitní nestabilita * MeSH
• mutace MeSH
• promotorové oblasti (genetika) MeSH
• regulace genové exprese u nádorů MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• staging nádorů MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Objev indukovaných pluripotentních kmenových buněk profesorem Shinya Yamanakem, který byl v roce 2012 oceněn Nobelovou cenou, otevřel nové horizonty v oblasti regenerativní medicíny, in vitro modelování nemocí nebo screeningu léčiv. Unikátní technologie, která umožňuje přípravu eticky přijatelných autologních kmenových buněk, činí reálnou oblast personalizované medicíny. Technologie iPSCs našla využití rovněž při in vitro modelování, kde umožnila přípravu buněk těžko dostupných tkání, např. neuronů nebo kardiocytů, z poměrně lehce dostupného zdroje, kterým jsou např. dermální fibroblasty. Takto připravené in vitro biomedicínské modely jsou aplikovatelné v preklinických studiích resp. při testování léčiv. Technologie iPSCs má široký potenciál využití v různých medicínských oblastech a rovněž již našla upotřebení také v boji proti nové pandemii koronaviru.

Discovery of induced pluripotent stem cells by Dr. Shinya Yamanaka, awarded by Nobel Prize in 2012 opened new horizonts in the field of regenerative medicine, in vitro disease modeling and drug screening. A unique technology that allows preparation of ethical issues-free, autologous therapeutic stem cells is revolutionizing the field of patient -tailored therapies. For in vitro modeling, iPSc technology represents a unique possibility for preparation of hard -to -get cells (neurons,cardiomyocytes etc) from patient skin fibroblasts for further studies and drug testing. iPSc technology has potential and is already used in world research effort in fight against new coronavirus pandemy.

The colorectal cancer harbor germline, somatic or epimutations in mismatch repair genes, MUTYH or POLE gene, which lead to the hypermutated and ultramutator phenotypes with increased immune response. The mutations in POLE gene were reported to occur more frequently in early-onset colorectal cancer (EOCRC), and the patients are strong candidates for checkpoint inhibitor therapy. Here, we report mutation analysis within the endonuclease domain of the POLE gene in the cohort of patients with EOCRC in order to identify recurrent or new mutations and evaluate their association with the presence of tumor-infiltrating lymphocytes (TILs) and peritumoral lymphoid reaction. We have shown a significant association between MSI tumors and TILs (p = 0.004). Using sensitive single-tube nested PCR with subsequent Sanger sequencing, we have found in one female patient diagnosed at age 48 with rectal adenocarcinoma with mucinous elements staged pT3pN2pM1 a silent variant within the exon 9 NM_006231.3 c.849 C > T, NP_00622.2 p.Leu283 = recorded in dSNP as rs1232888774 with MAF = 0.00002. In silico prediction, result showed possible involvement into splicing; therefore, this rare variant can be involved into EOCRC pathogenesis. In the time of precise medicine, it is important to develop screening strategies also for less common conditions such as EOCRC allowing to predict tailored therapy for younger patients suffering from CRC that harbor mutations in the POLE gene.

• MeSH
• DNA-polymerasa II genetika   MeSH
• dospělí MeSH
• genotypizační techniky MeSH
• kolorektální nádory genetika   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace * MeSH
• proteiny vázající poly-ADP-ribosu genetika   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The proto-oncogene KRAS belongs among the most frequently mutated genes in all types of cancer and is also very important oncogene related to colorectal tumors. The detection of mutations in this gene in primary tumor is a predictive biomarker for the anti-EGFR therapy in metastatic CRC (mCRC); however, the patients with wild-type KRAS can also show resistance to the personalized medicine. The droplet-based digital PCR technology has improved the analytical sensitivity of the mutations detection, which led us to the idea about the optimization of this approach for KRAS testing. In this study, we report the application of ddPCR technology in order to analyze the presence of KRAS mutations in primary tumor and matched metastasis in lymph nodes (LNs) from patients with mCRC and address the question, whether the improvement in the detection method can lower the discrepancies of KRAS mutations detection between the primary tumor and regional LNs. Genomic DNA with wtKRAS and commercial DNA with mtKRAS (G12D) were used to set up the ddPCR reaction. Formalin-fixed paraffin-embedded tissues from primary tumor and positive lymph node from 31 patients with mCRC were analyzed using ddPCR and Sanger sequencing. KRAS status of primary tumors was known; however, the mutation status of lymph nodes was not detected previously. From 31 samples of primary tumors, our results corresponded to results from IVD kit in 30 cases. For one patient, ddPCR detected KRAS mutation in comparison with negative result of the IVD kit. In the samples of metastatic infiltrated LNs, ddPCR detected 16 samples as a WT KRAS and 15 lymph nodes showed positivity for KRAS mutation, whereby Sanger sequencing found KRAS mutations in 8 cases only. We also found two cases where genetic conditions of KRAS gene differed between primary tumor and infiltrated lymph node, both "low-grade" adenocarcinoma. Our study approved that ddPCR method is adequate technique with high sensitivity and in the future may be used as a diagnostic tool for evaluation of KRAS mutations, especially in infiltrated LNs of patients with mCRC.

• MeSH
• kolon patologie   MeSH
• kolorektální nádory diagnóza   genetika   patologie   MeSH
• lidé MeSH
• lymfatické uzliny patologie   MeSH
• molekulární patologie metody   MeSH
• mutantní proteiny genetika   MeSH
• polymerázová řetězová reakce metody   MeSH
• protoonkogenní proteiny p21(ras) genetika   MeSH
• senioři MeSH
• senzitivita a specificita MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH

The development of the new technologies such as the next-generation sequencing (NGS) makes more accessible the diagnosis of genetically heterogeneous diseases such as Lynch syndrome (LS). LS is one of the most common hereditary form of colorectal cancer. This autosomal dominant inherited disorder is caused by deleterious germline mutations in one of the mismatch repair (MMR) genes - MLH1, MSH2, MSH6 or PMS2, or the deletion in the EPCAM gene. These mutations eventually result in microsatellite instability (MSI), which can be easily tested in tumor tissue. According to the actual recommendations, all patients with CRC that are suspect to have LS, should be offered the MSI testing. When the MSI is positive, these patients should be recommended to genetic counseling. Here we report a pilot study about the application of NGS in the LS diagnosis in patients considered to have sporadic colorectal cancer. The inclusion criteria for the NGS testing were MSI positivity, BRAF V600E and MHL1 methylation negativity. We have used 5 gene amplicon based massive parallel sequencing on MiSeq platform. In one patient, we have identified a new pathogenic mutation in the exon 4 of the MSH6 gene that was previously not described in ClinVar, Human Gene Mutation Database, Ensembl and InSight databases. This mutation was confirmed by the Sanger method. We have shown that the implementation of new criteria for colorectal patients screening are important in clinical praxis and the NGS gene panel testing is suitable for routine laboratory settings.

• MeSH
• dědičné nepolypózní kolorektální nádory diagnóza   genetika   MeSH
• lidé MeSH
• mikrosatelitní nestabilita MeSH
• oprava chybného párování bází DNA MeSH
• pilotní projekty MeSH
• vysoce účinné nukleotidové sekvenování * MeSH
• zárodečné mutace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Slovenská republika MeSH