Flow cytometry immunophenotyping is critical for the diagnostic classification of mature/peripheral B-cell neoplasms/B-cell chronic lymphoproliferative disorders (B-CLPD). Quantitative driven classification approaches applied to multiparameter flow cytometry immunophenotypic data can be used to extract maximum information from a multidimensional space created by individual parameters (e.g., immunophenotypic markers), for highly accurate and automated classification of individual patient (sample) data. Here, we developed and compared five diagnostic classification algorithms, based on a large set of EuroFlow multicentric flow cytometry data files from a cohort 659 B-CLPD patients. These included automatic population separators based on Principal Component Analysis (PCA), Canonical Variate Analysis (CVA), Neighbourhood Component Analysis (NCA), Support Vector Machine algorithms (SVM) and a variant of the CA(Canonical Analysis) algorithm, in which the number of SDs (Standard Deviations) varied for each of the comparisons of different pairs of diseases (CA-vSD). All five classification approaches are based on direct prospective interrogation of individual B-CLPD patients against the EuroFlow flow cytometry B-CLPD database composed of tumor B-cells of 659 individual patients stained in an identical way and classified a priori by the World Health Organization (WHO) criteria into nine diagnostic categories. Each classification approach was evaluated in parallel in terms of accuracy (% properly classified cases), precision (multiple or single diagnosis/case) and coverage (% cases with a proposed diagnosis). Overall, average rates of correct diagnosis (for the nine B-CLPD diagnostic entities) of between 58.9 % and 90.6 % were obtained with the five algorithms, with variable percentages of cases being either misclassified (4.1 %-14.0 %) or unclassifiable (0.3 %-37.0 %). Automatic population separators based on CA, SVM and PCA showed a high average level of correctness (90.6 %, 86.8 %, and 86.0 %, respectively). Nevertheless, this was at the expense of proposing a considerable number of multiple diagnoses for a significant proportion of the test cases (54.5 %, 53.5 %, and 49.6 %, respectively). The CA-vSD algorithm generated the smaller average misclassification rate (4.1 %), but with 37.0 % of cases for which no diagnosis was proposed. In contrast, the NCA algorithm left only 2.7 % of cases without an associated diagnosis but misclassified 14.0 %. Among correctly classified cases (83.3 % of total), 91.2 % had a single proposed diagnosis, 8.6 % had two possible diagnoses, and 0.2 % had three. We demonstrate that the proposed AI algorithms provide an acceptable level of accuracy for the diagnostic classification of B-CLPD patients and, in general, surpass other algorithms reported in the literature.

• MeSH
• Algorithms MeSH
• B-Lymphocytes * pathology   MeSH
• Immunophenotyping * methods   MeSH
• Middle Aged MeSH
• Humans MeSH
• Lymphoproliferative Disorders * diagnosis   classification   MeSH
• Flow Cytometry * methods   MeSH
• Aged MeSH
• Support Vector Machine MeSH
• Check Tag
• Middle Aged MeSH
• Humans MeSH
• Male MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Journal Article MeSH
• Comparative Study MeSH

Schizophrenia is a psychiatric disorder with heterogeneous clinical manifestations and complex aetiology. Notably, the triple-network model proposes an interesting framework for investigating abnormal neurocircuit activity at rest in schizophrenia. The present study on 30 chronic schizophrenia individuals and 30 controls aimed to explore the differences in EEG resting state effective connectivity within a triple-network model using source-localization-based Directed Transfer Function. Our findings revealed multiband effective connectivity disturbances within default mode (DMN), central executive (CEN), and salience (SN) networks in schizophrenia. The most significant difference was manifested in a global DMN hyperconnectivity, accompanied by low-band hyperconnectivity and high-band hypoconnectivity in CEN, along with the aberrant information flows in SN. In conclusion, our study presents novel insights into schizophrenia neuropathology, with a particular emphasis on the reversed directionality in information flows between hubs of SN, DMN, and CEN. This may be suggested as a promising biomarker of schizophrenia.

• MeSH
• Default Mode Network * physiopathology   MeSH
• Adult MeSH
• Electroencephalography MeSH
• Connectome * methods   MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Nerve Net * physiopathology   diagnostic imaging   MeSH
• Schizophrenia * physiopathology   diagnostic imaging   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

Postižení ledvin v rámci Light Chain Depositon Disease (LCDD) je velmi vzácně diagnostikovanou jednotkou. Popisuje- me případ, kdy tato diagnóza byla morfologicky stanovena až v biopsii transplantované ledviny a retrospektivně byla dohledána i v předchozí biopsii ledvin, kde však změny nebyly správně klasifikovány. Biopsie transplantované ledviny byla provedena pro postupně se horšící funkce štěpu. Následné vyšetření, cílené na monoklonální gamapatii, prokázalo zvýšenou sérovou koncentraci volných lehkých řetězců kappa (free light chain – FLC) s maximální hodnotou FLC kappa 226 mg/l a FLC lambda jen 6 mg/l. Poměr FLC kappa / FLC lambda byl jasně patologický, 37 (normalní rozmezí 0,26–1,65). Imunofixační elektroforéza séra a moče byla opakovaně negativní. Cytologické vyšetření kostní dřeně popsalo 8 % patolo- gických plazmatických buněk. Flow-cytometrické vyšetření kostní dřeně prokázalo 0,7 % plazmocytů ze všech jaderných buněk kostní dřeně. Tyto plazmocyty byly ve 100 % klonální, abnormálního fenotypu kappa+. Diagnóza byla uzavřena jako nemaligní gamapatie typu „monoklonální gamapatie renálního významu“ s poškozením ledvin v morfologické formě odpovídající LCDD. Pro léčbu byla zvolena kombinace daratumumabu, bortezomibu, cyklofosfamidu a dexametazonu. Současně pacientka dostávala imunosupresivní léčbu nutnou k zachování funkce transplantované ledviny. Sérová hladina volných lehkých řetězců kappa v průběhu prvních dvou měsíců léčby poklesla pod dolní hranici normy. LCDD je jednou z mnoha forem poškození ledvin, k němuž může dojít při nemaligních gamapatiích. Proto by vyšetření FLC mělo být provedeno vždy v rámci diferenciální diagnostiky každého renálního selhání. Pro poškození ledvin mono- klonálním imunoglobulinem byla akceptována klasifikace vytvořená mezinárodní skupinou The International Kidney and Monoclonal Gammopathy Research Group. Morfology, hodnoticí biopsie ledvin, je vhodné informovat o případné přítomnosti patologické koncentrace FLC anebo M-Ig, aby

Light Chain Deposition Disease (LCDD) is a very rarely diagnosed condition affecting the kidneys. We describe a case where this diagnosis was morphologically confirmed in a biopsy of a transplanted kidney, and retrospectively identified in a previous kidney biopsy where the changes were not correctly classified. The biopsy of the transplanted kidney was performed due to worsening graft function. Subsequent testing focused on monoclonal gammopathy, revealing elevated serum concentrations of free kappa light chains (FLC) with a maximum FLC kappa value of 226 mg/l and FLC lambda at only 6 mg/l. The FLC kappa / FLC lambda ratio was clearly pathological at 37 (normal range 0.26-1.65). Serum and urine immunofixation electrophoresis were repeatedly negative. Bone marrow cytology described 8% pathological plasma cells, and flow cytometry demonstrated 0.7% plasma cells among all nuclear bone marrow cells. These plasma cells were 100% clonal, of the abnormal kappa + phenotype. The diagnosis was thus concluded as a non-malignant gammopathy of the type "monoclonal gammopathy of clinical significance" with renal damage in a morphological form corresponding to LCDD. A combination of daratumumab, bortezomib, cyclophosphamide and dexamethasone was chosen for treatment LCDD. Free light kappa chains decreased below the lower limit of the norm during the first two months of anti-CD38 therapy. LCDD is one of the many forms of kidney damage that can occur in non-malignant gammopathies. Therefore, FLC testing should always be performed as part of the differential diagnosis of renal failure. For kidney damage by monoclonal immunoglobulin, a classification created by The International Kidney and Monoclonal Gammopathy Research Group was accepted. It is advisable to inform the evaluating morphologists of the possible presence of pathological concentrations of FLC and/or M-Ig so that they can focus the diagnosis in this direction, otherwise these rare forms of kidney injury may remain unrecognized.

• Keywords
• daratumumab,
• MeSH
• Bortezomib administration & dosage   therapeutic use   MeSH
• Cyclophosphamide administration & dosage   therapeutic use   MeSH
• Dexamethasone administration & dosage   therapeutic use   MeSH
• Immunoglobulin Light Chains blood   MeSH
• Humans MeSH
• Antibodies, Monoclonal administration & dosage   therapeutic use   MeSH
• Paraproteinemias diagnosis   pathology   therapy   MeSH
• Immunoglobulin Light-chain Amyloidosis * diagnosis   drug therapy   MeSH
• Renal Insufficiency etiology   MeSH
• Aged MeSH
• Kidney Transplantation MeSH
• Check Tag
• Humans MeSH
• Aged MeSH
• Female MeSH
• Publication type
• Case Reports MeSH

During hypoxia, tissues are subjected to an inadequate oxygen supply, disrupting the balance needed to maintain normal function. This deficiency can occur due to reduced oxygen delivery caused by impaired blood flow or a decline in the blood's ability to carry oxygen. In tumors, hypoxia and vascularization play crucial roles, shaping their microenvironments and influencing cancer progression, response to treatment and metastatic potential. This chapter provides guidance on the use of non-invasive imaging methods including Positron Emission Tomography and Magnetic Resonance Imaging to study tumor oxygenation in pre-clinical settings. These imaging techniques offer valuable insights into tumor vascularity and oxygen levels, aiding in understanding tumor behavior and treatment effects. For example, PET imaging uses tracers such as [18F]-fluoromisonidazole (FMISO) to visualize hypoxic areas within tumors, while MRI complements this with anatomical and functional images. Although directly assessing tumor hypoxia with MRI remains challenging, techniques like Blood Oxygen Level Dependent (BOLD) and Dynamic Contrast-Enhanced MRI (DCE-MRI) provide valuable information. BOLD can track changes in oxygen levels during oxygen challenges, while DCE-MRI offers real-time access to perfusion and vessel permeability data. Integrating data from these imaging modalities can help assess oxygen supply, refine treatment strategies, enhance therapeutic effectiveness, and ultimately improve patient outcomes.

• MeSH
• Hypoxia diagnostic imaging   MeSH
• Oxygen metabolism   MeSH
• Humans MeSH
• Magnetic Resonance Imaging * methods   MeSH
• Misonidazole analogs & derivatives   MeSH
• Mice MeSH
• Tumor Hypoxia MeSH
• Neoplasms diagnostic imaging   blood supply   pathology   MeSH
• Neovascularization, Pathologic diagnostic imaging   pathology   MeSH
• Positron-Emission Tomography * methods   MeSH
• Animals MeSH
• Check Tag
• Humans MeSH
• Mice MeSH
• Animals MeSH
• Publication type
• Journal Article MeSH

Frekvence familiární hypercholesterolémie (FH) je u většiny populací asi 1/200 a je tedy možné předpokládat, že v ČR by mohlo být postiženo přibližně 50 000 osob. Řešení projektu lze rozdělit do dvou pracovních bloků (PB). PB1 je zaměřen na funkční analýzu variant genu LDLR in vitro a využívá metod průtokové cytometrie, konfokální mikroskopie na živých buňkách, qRT-PCR a sestřihových minigenových testů. Varianty LDLR budou hodnoceny na úrovni exprese, zrání, lokalizace a funkce daného proteinu a na úrovni buněčné odpovědi (aktivace tzv. unfolded protein response). PB2 je zaměřen na analýzu DNA pacientů. Metodou NGS bude provedena re-analýza pacientů s FH s dosud negativní genetickou diagnózou a budou aplikovány metody pro stanovení počtu repetic Kringle IV v genu LPA (PFGE a Southernova hybridizace) pro rozřazení pacientů do skupiny s nižšími nebo vyšší riziky předčasné aterosklerózy. Všechny uvedené přístupy mají společný cíl – identifikovat genetické varianty způsobující FH a získat nové informace o patogenitě FH.; The frequency of familial hypercholesterolemia (FH) in most populations is about 1/200, and so it is possible to predict that about 50,000 people could be affected by FH in the Czech Republic. The project solution can be divided into two work packages (WP). WP1 is focused on functional analysis of LDLR variants in vitro and methods as flow cytometry, live cell imaging microscopy, qRT-PCR, splicing minigene assay will be applied. LDLR variants will be evaluated on level of protein expression, maturation, localisation, function, and cell response (activation of the unfolded protein response). WP2 is focused on analysis of patients ́ DNA. Re-analysis of FH patients with so far negative genetic diagnosis will be performed by NGS, and methods for determination of the Kringle IV repeat number in the LPA gene (PFGE and Southern hybridisation) will be applied for stratification of patients into a group with lower or higher risk of premature atherosclerosis. All mentioned approaches have a common goal – to identify FH disease-causing variants and obtain new information about FH pathogenicity.

• Keywords
• familial hypercholesterolemia, Familiální hypercholesterolémie, funkční analýza, functional analysis, confocal microscopy, konfokální mikroskopie, LDLR, LDLR, skládání proteinů, lipoprotein(a), protein folding, lipoprotein(a),

• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

Nové metody jako sekvenování nové generace (NGS) přinesly obrovské množství informací o nádorové biologii a umožnily vývoj nových metod pro diagnostiku a sledování zbytkové choroby (MRN). Transfer nových metod do praxe je komplikován jejich množstvím, finanční náročností i redundancí. V mezinárodní spolupráci chceme vyvinout nástroj na simultánní detekci klonality, MRN a profilu přestaveb genů pro imunoglobuliny a T-buněčné receptory infiltrujících fyziologických lymfocytů. Postup bude sjednocovat a zjednodušovat stávající metodiky a zároveň poskytne nová data o imunitní odpovědi na nádor. Metodu chceme validovat na retrospektivní kohortě pacientů s anaplastickým velkobuněčným lymfomem a prospektivně na vzorcích dětských a adolescentních pacientů s lymfomy. Využití u jiných než lymfoidních malignit chceme testovat u dětí s mozkovými tumory (low- a high-grade gliomy). Výstupem projektu bude kromě standardizovaného metodického postupu prohloubení znalostí o složení a funkci infiltrujících imunitních buněk (především T lymfocytů) a zmapování prostoru pro cílení imunoterapie.; Next generation sequencing (NGS) and other new methods brought a large amount of data on tumour biology and facilitated the development of new methods for diagnostics and minimal residual disease (MRD) detection. The transfer of those methods into routine management is complicated by their numbers, costs and redundancy. In international cooperation we want to develop a tool for simultaneous detection of clonality, MRD and profile of immunoglobulin and T-cell receptor gene rearrangements in infiltrating physiological lymphocytes. The methodology will unite and simplify the current methods, while providing new data on immune response. We will validate the method on retrospective cohort of patients with anaplastic large cell lymphoma and prospectively on samples from children and adolescents with lymphoma. The utility in non-lymphoid malignancies will be tested in children with low-and high-grade gliomas. Next to the standardized methodology, the project will bring new information on the composition of infiltrating immune cells (mainly T lymphocytes) and a potential for immunotherapy.

• Keywords
• sekvenování nové generace, průtoková cytometrie, minimální reziduální nemoc, minimal residual disease, flow cytometry, lymfom, lymphoma, Next-generation sequencing, immune monitoring, cell-free DNA, tumor infiltrující lymfocyty, Immune checkpoints, imunomonitoring, imunitní kontrolní body, přestavby genů pro imunoglobuliny a T-buněčné receptory, gamadelta T lymfocyty, immunoglobulin and T-cell rearrangement, gammadelta T lymphocytes, volná nádorová DNA, tumour infiltrating lymphocytes,

• NML Publication type
• závěrečné zprávy o řešení grantu AZV MZ ČR

Antagonistic activity of brain networks likely plays a fundamental role in how the brain optimizes its performance by efficient allocation of computational resources. A prominent example involves externally/internally oriented attention tasks, implicating two anticorrelated, intrinsic brain networks: the default mode network (DMN) and the dorsal attention network (DAN). To elucidate electrophysiological underpinnings and causal interplay during attention switching, we recorded intracranial EEG (iEEG) from 25 epilepsy patients with electrode contacts localized in the DMN and DAN. We show antagonistic network dynamics of activation-related changes in high-frequency (> 50 Hz) and low-frequency (< 30 Hz) power. The temporal profile of information flow between the networks estimated by functional connectivity suggests that the activated network inhibits the other one, gating its activity by increasing the amplitude of the low-frequency oscillations. Insights about inter-network communication may have profound implications for various brain disorders in which these dynamics are compromised.

• MeSH
• Adult MeSH
• Electroencephalography MeSH
• Electrophysiological Phenomena MeSH
• Epilepsy physiopathology   MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Brain * physiology   physiopathology   MeSH
• Nerve Net * physiology   MeSH
• Attention * physiology   MeSH
• Check Tag
• Adult MeSH
• Middle Aged MeSH
• Humans MeSH
• Young Adult MeSH
• Male MeSH
• Female MeSH
• Publication type
• Journal Article MeSH

BACKGROUND: Understanding the risk factors leading to intracranial aneurysm (IA) rupture have still not been fully clarified. They are vital for proper medical guidance of patients harboring unruptured IAs. Clarifying the hemodynamics associated with the point of rupture could help could provide useful information about some of the risk factors. Thus far, few studies have studied this issue with often diverging conclusions. METHODS: We identified a point of rupture in patients operated for an IAs during surgery, using a combination of preoperative computed tomography (CT) and computed tomography angiography (CTA). Hemodynamic parameters were calculated both for the aneurysm sac as a whole and the point of rupture. In two cases, the results of CFD were compared with those of the experiment using particle image velocimetry (PIV). RESULTS: We were able to identify 6 aneurysms with a well-demarcated point of rupture. In four aneurysms, the rupture point was near the vortex with low wall shear stress (WSS) and high oscillatory shear index (OSI). In one case, the rupture point was in the flow jet with high WSS. In the last case, the rupture point was in the significant bleb and no specific hemodynamic parameters were found. The CFD results were verified in the PIV part of the study. CONCLUSION: Our study shows that different hemodynamic scenarios are associated with the site of IA rupture. The numerical simulations were confirmed by laboratory models. This study further supports the hypothesis that various pathological pathways may lead to aneurysm wall damage resulting in its rupture.

• Publication type
• Journal Article MeSH

Pulmonary hypertension (PH) associated with left heart failure (LHF) (PH-LHF) is one of the most common causes of PH. It directly contributes to symptoms and reduced functional capacity and negatively affects right heart function, ultimately leading to a poor prognosis. There are no specific treatments for PH-LHF, despite the high number of drugs tested so far. This scientific document addresses the main knowledge gaps in PH-LHF with emphasis on pathophysiology and clinical trials. Key identified issues include better understanding of the role of pulmonary venous versus arteriolar remodelling, multidimensional phenotyping to recognize patient subgroups positioned to respond to different therapies, and conduct of rigorous pre-clinical studies combining small and large animal models. Advancements in these areas are expected to better inform the design of clinical trials and extend treatment options beyond those effective in pulmonary arterial hypertension. Enrichment strategies, endpoint assessments, and thorough haemodynamic studies, both at rest and during exercise, are proposed to play primary roles to optimize early-stage development of candidate therapies for PH-LHF.

• MeSH
• Ventricular Function, Right * physiology   MeSH
• Humans MeSH
• Hypertension, Pulmonary * physiopathology   etiology   therapy   MeSH
• Pulmonary Circulation * physiology   MeSH
• Heart Failure * physiopathology   complications   therapy   MeSH
• Check Tag
• Humans MeSH
• Publication type
• Journal Article MeSH
• Review MeSH

Nepředpokládáme, že by pan profesor Barek páchal trestnou činnost, ale pokud bychom jeho pachovou stopu přesto odebrali a provedli detailní chemickou analýzu, mohli bychom se přesvědčit o tom, že je to muž, že je europoidní rasy a případně zjistit, jaká v jeho žilách koluje krev, zda patří do skupiny A, B nebo 0. Pachová stopa je zdroj informací, který orgány činné v trestním řízení prostřednictvím speciálně cvičených psů používají pro individuální identifikaci již desítky let. Nyní je snaha vytvořit instrumentální metodu pro detailní chemickou analýzu pachové stopy tak, aby pachová identifikace mohla být objektivizována a používána jako důkazní prostředek a samozřejmě, aby poskytovala řadu dalších informací o poskytovateli pachu.

We do not presume that professor Barek has committed any criminal act, but, if we nevertheless collect his scent sample and conduct a detailed chemical analysis, we could confirm that he is male, of the Caucasian race, and perhaps determine which blood type flows in his veins, whether it is A, B, or 0. Scent evidence is a source of information that prosecuting authorities have been using for decades to identify individuals through specially trained canines. There is now an effort to create an instrumental method for a detailed chemical analysis of the scent trace so that scent identification can be objectified and used as an evidence in court, and of course, to provide a range of additional pieces of information about the scent provider.

• Keywords
• olfaktronika,
• MeSH
• Biomarkers analysis   MeSH
• Chromatography, Gas MeSH
• Humans MeSH
• Odorants * analysis   MeSH
• Forensic Sciences MeSH
• Body Odor MeSH
• Check Tag
• Humans MeSH