The application of digital pathology and artificial intelligence in anatomical pathology represents a revolutionary step towards the modernization of diagnostic processes. Digitalization, primarily based on creation and subsequent use of whole slide imaging, enables generating of full digital images of histological slides, offering potential benefits in diagnostic accuracy and accessibility. Unlike traditional microscopy, digital pathology also facilitates telemedicine and remote consultation, opening new possibilities for collaboration and sharing of expertise at both national and international levels. However, implementing a digital workflow requires substantial investments in scanners, software platforms, high-capacity storage, and IT infrastructure. Despite considerable costs of implementation, it brings numerous advantages, including time savings, opportunities for centralized diagnostics, and a reduction in sample transport costs. This paper focuses on the practical aspects of implementing digital pathology in pathology laboratories, emphasizing the benefits, risks, and technological requirements associated with digitalized workflows. It also discusses crucial roles of validation and verification, which are essential for ensuring a diagnostic accuracy of digital images compared to conventional microscopy. The article presents digital pathology as a dynamically evolving field with high potential for personalized medicine, improved diagnostic accuracy, and support for remote collaboration, addressing the growing demands of modern medicine.

• Klíčová slova
• Digital Pathology, artificial intelligence, machine learning, whole slide image,
• MeSH
• laboratorní medicína * metody   MeSH
• lidé MeSH
• mikroskopie MeSH
• počítačové zpracování obrazu * MeSH
• průběh práce * MeSH
• telepatologie MeSH
• umělá inteligence MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Gene sequencing of 16S, 18S, and ITS regions is a crucial tool in molecular diagnostics, especially in microbiology, pathology and forensic medicine. These genes contain conserved and variable regions and are widely used for the taxonomic classification of bacteria and eukaryotes. Sequencing of 16S rDNA helps detect bacterial infections, while sequencing of ITS regions and 18S rDNA is used to identify fungal or parasitic infections, especially when traditional methods are ineffective. This article focuses on the expanded possibilities of these methods, their application in clinical diagnostics and research, their advantages and disadvantages, and discusses potential future developments in the field of next-generation sequencing (NGS) technology.

• Klíčová slova
• 16S sequencing, 18S sequencing, ITS region sequencing, Infection diagnostics, Molecular pathology, Next-generation sequencing, next generation sequencing,
• MeSH
• diagnostické techniky molekulární MeSH
• lidé MeSH
• RNA ribozomální 16S * genetika   MeSH
• RNA ribozomální 18S * genetika   MeSH
• sekvenční analýza DNA MeSH
• vysoce účinné nukleotidové sekvenování MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• RNA ribozomální 16S * MeSH
• RNA ribozomální 18S * MeSH

Subependymal giant cell astrocytoma (SEGA) is a World Health Organization Central Nervous System grade 1 tumor, strongly associated with tuberous sclerosis complex (TSC). Recent research indicates that Glycoprotein Nonmetastatic Melanoma Protein B (GPNMB), regulated by microphthalmia (MiT) family transcription factors may also be modulated by loss-of-function mutations in TSC1/2. We evaluated GPNMB as a diagnostic marker of subependymal giant cell astrocytoma (SEGA). A total of 11 patients with SEGA were included in the study. The control group comprised 185 primary central nervous system tumors, including high-grade and low-grade gliomas and glioneuronal/neuronal tumors. Strong and diffuse (≥ 50% of tumor cells) GPNMB expression was present in all SEGAs. In contrast, TTF-1 expression was detected in nine SEGAs, resulting in a sensitivity of 81.8%. Among the control group, 77 cases (41.6%) were negative for GPNMB and 102 (55.1%) cases were scored as > 1% < 50% positive. Only six control tissues (3.2%) showed diffuse and strong GPNMB expression. Among the tumors with strong GPNMB expression, there were three glioblastomas (GBMs) with morphology potentially mimicking SEGA but lacking TSC1, TSC2, or MTOR mutations. Using a cutoff of diffuse (≥ 50%) and strong positivity, GPNMB demonstrated 100% sensitivity (95% confidence interval: 74.1%-100%) and 96.8% specificity (95% confidence interval: 93.1%-98.5%) for diagnosing SEGA.

• Klíčová slova
• GPNMB, Immunohistochemistry, Subependymal giant cell astrocytoma, TSC1/2, Tuberous sclerosis,
• Publikační typ
• časopisecké články MeSH

BACKGROUND AND OBJECTIVES: Disparities between tumors arising via different sporadic carcinogenetic pathways have not been studied systematically. This retrospective multicenter cohort study evaluated the differences in the risk for non-colorectal malignancy between sporadic colorectal cancer (CRC) patients from different DNA mismatch repair status. METHODS: A retrospective European multicenter cohort study including in total of 1706 CRC patients treated between 1996 and 2019 in three different countries. The proficiency (pMMR) or deficiency (dMMR) of mismatch repair was determined by immunohistochemistry. Cases were analyzed for tumor BRAFV600E mutation, and BRAF mutated tumors were further analyzed for hypermethylation status in the promoter region of MLH1 to distinguish between sporadic and hereditary cases. Swedish and Finish patients were matched with their respective National Cancer Registries. For the Czech cohort, thorough scrutiny of medical files was performed to identify any non-colorectal malignancy within 20 years before or after the diagnosis of CRC. Poisson regression analysis was performed to identify the incidence rates of non-colorectal malignancies. For validation purposes, standardized incidence ratios were calculated for the Swedish cases adjusted for age, year, and sex. RESULTS: Of the 1706 CRC patients included in the analysis, 819 were female [48%], median age at surgery was 67 years [interquartile range: 60-75], and sporadic dMMR was found in 188 patients (11%). Patients with sporadic dMMR CRC had a higher incidence rate ratio (IRR) for non-colorectal malignancy before and after diagnosis compared to patients with a pMMR tumor, in both uni- (IRR = 2.49, 95% confidence interval [CI] = 1.89-3.31, p = 0.003) and multivariable analysis (IRR = 2.24, 95% CI = 1.67-3.01, p = 0.004). This association applied whether or not the non-colorectal tumor developed before or after the diagnosis of CRC in both uni- (IRR = 1.91, 95% CI = 1.28-2.98, p = 0.004), (IRR = 2.45, 95% CI = 1.72-3.49, p = 0.004) and multivariable analysis (IRR = 1.67,95% CI = 1.05-2.65, p = 0.029), (IRR = 2.35, 95% CI = 1.63-3.42, p = 0.005), respectively. CONCLUSION: In this retrospective European multicenter cohort study, patients with sporadic dMMR CRC had a higher risk for non-colorectal malignancy than those with pMMR CRC. These findings indicate the need for further studies to establish the need for and design of surveillance strategies for patients with dMMR CRC.

• Klíčová slova
• colorectal cancer, non‐colorectal malignancy, sporadic deficient mismatch repair,
• MeSH
• incidence MeSH
• kolorektální nádory * genetika   patologie   epidemiologie   etiologie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mutace MeSH
• MutL homolog 1 genetika   MeSH
• následné studie MeSH
• oprava chybného párování bází DNA * MeSH
• prognóza MeSH
• protoonkogenní proteiny B-Raf genetika   MeSH
• retrospektivní studie MeSH
• senioři MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH
• Geografické názvy
• Evropa epidemiologie   MeSH
• Švédsko epidemiologie   MeSH
• Názvy látek
• BRAF protein, human MeSH Prohlížeč
• MLH1 protein, human MeSH Prohlížeč
• MutL homolog 1 MeSH
• protoonkogenní proteiny B-Raf MeSH

Traditionally considered non-functional low proliferative benign neuroendocrine proliferations measuring less than 5 mm, pancreatic (neuro)endocrine microadenomas are now classified as pancreatic neuroendocrine microtumors in the 2022 WHO classification of endocrine and neuroendocrine tumors. This case report discussed the features of an incidentally identified 4.7-mm glucagon-expressing pancreatic neuroendocrine microtumor with MEN1 mutation only, chromosomally stable and an epigenetic alpha-like phenotype. The tumor was associated with an unexplained increased proliferation rate in Ki-67 of 15%. There was no associated DAXX/ATRX deficiency. The presented case challenges the conventional thought of a low proliferative disease of the so-called "pancreatic neuroendocrine microadenomas" and provides additional support to the 2022 WHO classification that also requires grading of these neoplasms. Despite exhibiting molecular features of less aggressive behavior, the case also underscores the biological complexity of pancreatic neuroendocrine microtumors. By recognizing the heterogenous spectrum of neuroendocrine neoplasms, the current case also contributes to ongoing discussions on how to optimize the clinical management of such tumors.

• Klíčová slova
• Ki-67, MEN1, Microadenoma, Neuroendocrine neoplasm, PanNETs, Pancreatic neuroendocrine microtumors, WHO 2022,
• MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory slinivky břišní * patologie   genetika   MeSH
• neuroendokrinní nádory * patologie   genetika   MeSH
• proliferace buněk MeSH
• protoonkogenní proteiny genetika   MeSH
• stupeň nádoru MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Názvy látek
• MEN1 protein, human MeSH Prohlížeč
• protoonkogenní proteiny MeSH

Low-grade serous carcinoma (LGSC) may develop from serous borderline tumor (SBT) tissue, where the micropapillary type (mSBT) presents the highest risk for progression. The sensitivity of LGSC to standard chemotherapy is limited, so alternative therapeutic approaches, including targeted treatment, are needed. However, knowledge about the molecular landscape of LGSC and mSBT is limited. A sample set of 137 pathologically well-defined cases (LGSC, 97; mSBT, 40) was analyzed using capture DNA next-generation sequencing (727 genes) and RNA next-generation sequencing (147 genes) to show the landscape of somatic mutations, gene fusions, expression pattern, and prognostic and predictive relevance. Class 4/5 mutations in the main driver genes (KRAS, BRAF, NRAS, ERBB2, USP9X) were detected in 48% (14/29) of mSBT cases and 63% (47/75) of LGSC cases. The USP9X mutation was detected in only 17% of LGSC cases. RNA next-generation sequencing revealed gene fusions in 6 of 64 LGSC cases (9%) and 2 of 33 mSBT cases (9%), and a heterogeneous expression profile across LGSC and mSBT. No molecular characteristics were associated with greater survival. The somatic genomic and transcriptomic profiles of 35 mSBT and 85 LGSC cases are compared for the first time. Candidate oncogenic gene fusions involving BRAF, FGFR2, or NF1 as a fusion partner were identified. Molecular testing of LGSC may be used in clinical practice to reveal therapeutically significant targets.

• MeSH
• azosloučeniny * MeSH
• genomika MeSH
• lidé MeSH
• mutace MeSH
• nádory vaječníků * diagnóza   genetika   MeSH
• protoonkogenní proteiny B-Raf genetika   MeSH
• RNA MeSH
• serózní cystadenokarcinom * diagnóza   genetika   MeSH
• stanovení celkové genové exprese MeSH
• stupeň nádoru MeSH
• thiolesterasa ubikvitinu genetika   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• azosloučeniny * MeSH
• Martius scarlet blue trichrome MeSH Prohlížeč
• protoonkogenní proteiny B-Raf MeSH
• RNA MeSH
• thiolesterasa ubikvitinu MeSH
• USP9X protein, human MeSH Prohlížeč

The current progress and increasing knowledge about the genetic causes of cancer opens up new possibilities for its treatment. However, it is necessary to combine the results obtained using classical pathological methods with sensitive, multiplex molecular pathological methods. The method that meets the required criteria is MLPA based on multiplex PCR reaction. This method detects both changes in gene copy number and DNA methylation and, last but not least, point mutations. The MLPA reaction is applicable to even highly fragmented DNA. At the same time, it is a robust method that can be performed on standard thermocyclers, the fluorescent tip label requires automatic sequencers. Up to 50 genetic markers can be tested in one reaction, a number that allows a diagnostic and prognostic conclusion. All these features lead to the routine use of MLPA analysis not only in diagnosis but also in cancer research. The present article aims to summarize the different types of MLPA reactions, its benefits, but also the potential pitfalls.

• Klíčová slova
• MLPA, Molecular pathology, PCR, Tumors of the central nervous system,
• MeSH
• DNA genetika   MeSH
• genetické markery MeSH
• genová dávka MeSH
• lidé MeSH
• metylace DNA * MeSH
• nádory centrálního nervového systému * genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• DNA MeSH
• genetické markery MeSH

The aim of the presented communication is to clearly inform the general professional public about the newly approved modifications in this classification, including the newly approved types of tumours. A significant change is the new grading system for these tumours, including the innovative involvement of tumour profiling at the molecular level in the system for determining the degree of tumour differentiation and the application of the principle of integrated diagnostics, i. e. the synthesis of available histopathological and molecular findings in CNS tumors.

• Klíčová slova
• Molecular pathology, WHO classification, grading, tumours of the central nervous system,
• MeSH
• lidé MeSH
• nádory centrálního nervového systému * diagnóza   MeSH
• Světová zdravotnická organizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

INTRODUCTION: Radiation-associated angiosarcoma (RAAS) is a rare and serious complication of breast irradiation. Due to the rarity of the condition, clinical experience is limited and publications on this topic include only retrospective studies or case reports. MATERIALS AND METHODS: All patients diagnosed with RAAS between January 2000 and December 2017 in twelve centers across the Czech Republic and Slovakia were evaluated. RESULTS: Data of 53 patients were analyzed. The median age at diagnosis was 72 (range 44-89) years. The median latency period between irradiation and diagnosis of RAAS was 78 (range 36-172) months. The median radiation dose was 57.6 (range 34-66) Gy. The whole breast radiation therapy with radiation boost to the tumor bed was the most common radiotherapy regimen. Total mastectomy due to RAAS was performed in 43 patients (81%), radical excision in 8 (15%); 2 patients were not surgically treated due to unresectable disease. Adjuvant chemotherapy followed surgical therapy of RAAS in 18 patients, 3 patients underwent adjuvant radiotherapy. The local recurrence rate of RAAS was 43% and the median time from surgery to the onset of recurrence was 7.5 months (range 3-66 months). The 3-year survival rate was 56%, the 5-year survival rate was only 33%. 46% of patients died during the follow-up period. CONCLUSION: The present data demonstrate that RAAS is a rare condition with high local recurrence rate (43%) and mortality (the 5-year survival rate was 33%.). Early diagnosis of RAAS based on biopsy is crucial for treatment with radical intent. Surgery with negative margins constitutes the most important part of the therapy; the role of adjuvant chemotherapy and radiotherapy is still unclear.

• Klíčová slova
• Angiosarcoma, Breast, Multicenter study, Radiation,
• MeSH
• adjuvantní radioterapie * škodlivé účinky   MeSH
• dospělí MeSH
• hemangiosarkom * radioterapie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• lokální recidiva nádoru MeSH
• mastektomie MeSH
• nádory prsu * radioterapie   MeSH
• nádory vyvolané zářením * epidemiologie   MeSH
• následné studie MeSH
• retrospektivní studie MeSH
• segmentální mastektomie MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• multicentrická studie MeSH

Shortly after the WHOs first notice a suspected case of omicron SARS-CoV-2 was reported in Liberec, Czech Republic. The primary goal of the following actions was to test the presence of the variant and stop the spread of the virus variant. On November 25 a sixty-year-old lady, who had recently returned from Namibia, visited a GP with flu-like symptoms and a rash on her chest. The antigen test was positive for SARS-CoV-2, a PCR test was planned. At that time, it was not known that a new variant of concern was spreading from Africa. On November 26 in the morning the GP announced a suspected omicron case to the Regional public health authority, who organized the following steps. A mobile sampling team was sent to the patient's home immediately, sample transported into the regional hospital and analyzed with the help of the national reference laboratory. The captured virus SARS-CoV-2 fitted the description of the omicron variant, was shared in the GISAID database and named hCoV-19/Czech Republic/KNL_2021-110119140/2021. Contact tracing was started immediately, eleven persons were tested and quarantined. One of them positive with no further spread. It is the first documented omicron case in the Czech Republic and one of the first cases in Europe, with an excellent systemic response to the alert. The laboratory was able to detect the omicron variant instantly after the request. This case also demonstrates how easily the virus spreads on long distances and how important it might be to increase the uptake of the booster vaccine.

• Klíčová slova
• COVID-19, SARS-CoV-2, case report, omicron,
• MeSH
• COVID-19 * MeSH
• lidé středního věku MeSH
• lidé MeSH
• SARS-CoV-2 * MeSH
• výzkum MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH
• Geografické názvy
• Evropa MeSH