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A diffúz nagy B-sejtes limfóma 5 éves általános túlélése a mai kezelések mellett 60–70%, melynek hátterében a betegség komplex heterogenitása állhat.CélkitűzésMagyarországi multicentrikus tanulmányban a betegség fenotipikus, citogenetikai, genomexpressziós profil- és geográfiai heterogenitásának vizsgálata.MódszerA 276 formol-paraffinos betegmintát Hans' algoritmus és dupla protein expresszor státusz alapján klasszifikáltuk. A IGH::MYC, IGH::BCL2, BCL6 gén átrendeződéseket, valamint a MYC, BCL2 és BCL6 gének nyerését interfázis citogenetikával vizsgáltuk. Az RNA-seq-alapú génexpressziós profilvizsgálat 173 formol-paraffinos mintán volt elvégezhető.EredményekA Hans' fenotípus alapján 103 (37,3%) germinális centrum B-sejt-szerű és 173 (62,7%) nem germinalis B-sejt-szerű limfómát azonosítottunk, mely besorolás 82,6%-os megegyezést mutatott a genomexpressziós profilstratifikációval. Tripla aberrációt mutató limfóma nem fordult elő. Izolált IGH::MYC átrendeződés, valamint MYC, BCL2, BCL6 géntöbblet mindkét Hans' csoportban jelen volt. Az IGH::BCL2 átrendeződés izoláltan vagy kombinációban, szignifikánsan és kizárólag a germinális centrum B-sejt-szerű csoportban volt azonosítható, míg a BCL6 átrendeződés szignifikáns halmozódást mutatott a nem germinális centrum B-sejt-szerű Hans' csoportban. A dupla protein expresszor fenotípus pozitív prediktív értéke mindkét Hans' csoport molekuláris alcsoportjaira alacsony, a 0,04–0,19, illetve a 0,12–0,30, tartományba esett, míg negatív prediktív értéke mindkét főcsoport összes releváns molekuláris alcsoportjában 1,00-nek felelt meg.KövetkeztetésekA IGH::MYC átrendeződés nem Hans' csoport specifikus genotípus. Az IGH::BCL2 átrendeződés germinalis centrum, a BCL6 átrendeződés pedig a nem germinalis centrum B-sejt-szerű limfóma fémjele. A dupla protein expresszor negatív fenotípus mellett IGH::MYC és/vagy IGH::BCL2 átrendeződés nem fordult elő.

50 éves CML-es beteg esetét mutatjuk be, akinek a betegség diagnózisakor a Ph+ klón klonális evolúcióját (+8) észleltük. A második generációs TKI (nilotinib) kezelésre mély molekuláris remisszióba jutott betegben ez megszűnt, de a kezelés 30. hónapjában a Ph– klón evolúciója 5q- és 7-es kromoszómavesztéssel járó MDS kialakulását eredményezte. 8 hónapos masszív hemoszupportáció és lenalidomid kezelés mellett átmenetileg a Ph–, nem MDS klónból történő vérképzése visszatért és transzfúziót sem igényelt. Az allogén őssejt-transzplantációt visszautasította. 4 hónap tünetmentességet követően az MDS klón újra visszatért, és a beteg neutropeniás szepszisben meghalt.

Összefoglaló. A Magyar Hematológiai és Transzfuziológiai Társaság (MHTT) Onkohematológiai Molekuláris Diagnosztikai Munkacsoportja öt egyetemi és országos onkohematológiai centrumban 2018 és 2020 között 2261 krónikus limfocitás leukémiában (CLL) szenvedő beteg immunglobulin nehézláncgénjének (IGHV-) vizsgálatát végezte el a rutindiagnosztika keretében, jelen írásunkban ezen eredményeket foglaljuk össze. A vizsgált minták 53,5%-a bizonyult nem mutált immunglobulin nehézlánccal rendelkező CLL-nek, mutált immunglobulint az esetek 41,6%-ában detektáltunk. Az átrendeződött immunglobulin-szekvenciákat 11%-ban tudtuk valamelyik CLL alcsoportba („subset”) besorolni, melyek közül a CLL#2, a CLL#1, a CLL#6, CLL#4, CLL#3 és a CLL#5 alcsoport bizonyult a leggyakoribbnak. A szerzők kiemelik, hogy az elmúlt évek fejlesztéseinek köszönhetően valamennyi hazai onkohematológiai diagnosztikus laboratóriumban elérhető a nemzetközi ajánlások alapján kivitelezett IGHV-mutáció-analízis. Summary. The Hungarian Society of Hematology and Transfusiology’s Molecular Oncohematological Diagnostic Workgroup analyzed the mutational status of the immunoglobulin gene heavy chain region in 2261 CLL patient between 2018 and 2020 in five Hungarian oncohematological center. In this cohort 53.5% of the cases proved to have unmutated immunoglobulin gene, meanwhile mutated cases represented 41.6% of the entire cohort. We detected stereotype subsets in 11%, of which CLL#2, CLL#1, CLL#6, CLL#4, CLL#3, and CLL#5 were the most common. The authors highlight that collaborative efforts during the last few years lead to availability of the IGHV mutation analysis in the Hungarian diagnostic oncohematology laboratories as recommended by international standards.

OBJECTIVES: Esophageal pain is mediated by sensory nerves, most importantly via the activation of the Transient Receptor Potential Vanilloid 1 (TRPV1) capsaicin receptor. TRPV1 is activated and sensitized by a broad range of pungent compounds, as well as inflammatory mediators and tissue irritants. Luminal stressors are suggested to impair the barrier function, which results in consequent activation of these sensory nerve terminals and pain. Here we investigated the effect of the perfusion of capsaicin, a TRPV1 agonist, on mucosal impedance and pain in asymptomatic volunteers.METHODS: Thirteen asymptomatic volunteers completed a single blind, saline-controlled, randomized crossover study. Capsaicin or saline was perfused for 30 min in the distal oesophagus. Visual Analogue Scale (VAS) pain intensity scores and intraluminal impedance indicating mucosal integrity were determined. Distal and proximal biopsies were obtained 10 min later to measure TRPV1 mRNA and Trpv1 immunopositivity, as well as the intercellular space area.RESULTS: Capsaicin perfusion resulted in significantly greater pain intensity (P=0.047) and impaired recovery of the mucosal impedance compared to saline-treated controls (P=0.027). Pain response was significantly associated with decreased mucosal impedance. Similar dynamics were seen in the proximal esophagus, but mucosal impedance recovered entirely to the pre-infusion values there. There was a significant association between mucosal impedance and intercellular space width in the distal esophagus. TRPV1 transcription and expression were not significantly altered within this observation period.DISCUSSION: Esophageal capsaicin perfusion results in pain, which is likely to be explained by impaired mucosal impedance and defective restoration capacity in the distal esophagus.

In spite of intensive research, the survival rates of patients diagnosed with tumors of the central nervous system (CNS) have not improved significantly in the last decade. Immunotherapy as novel and efficacious treatment option in several other malignancies has failed in neuro-oncology likely due to the immunosuppressive property of the brain tissues. Glioblastoma (GBM) is the most aggressive malignant CNS neoplasm, while meningioma (MNG) is a mainly low grade or benign brain tumor originating from the non-glial tissues of the CNS. The aim of the current preliminary study is to compare the immune microenvironment of MNG and GBM as potential target in immunotherapy. Interestingly, the immune microenvironment of MNG and GBM have proved to be similar. In both tumors types the immune suppressive elements including regulatory T cells (Treg), tumor-associated macrophages (TAM) were highly elevated. The cytokine environment supporting Treg differentiation and the presence of indoleamine 2,3-dioxygenase 1 (IDO1) have also increased the immunosuppressive microenvironment. The results of the present study show an immune suppressive microenvironment in both brain tumor types. In a follow-up study with a larger patient cohort can provide detailed background information on the immune status of individual patients and aid selection of the best immune checkpoint inhibitor or other immune modulatory therapy. Immune modulatory treatments in combination with IDO1 inhibitors might even become alternative therapy for relapsed, multiple and/or malignant MNG or chemo-resistant GBM.

A case of a 61-year-old male patient suffered chronic renal failure and dialysed for 23 years with destructive cervical spondylarthropathy is presented. The patient presented with sudden onset of cervical pain radiating into his shoulders without neurological deficits. CT and MRI of the cervical and thoracic spine revealed severe destructive changes and compressive fractures of C6 and C7 vertebrae which caused the narrowing of the nerve root canals at these levels. A 360-degree fixation was performed to treat the unstable fracture and the patient’s pain (C6 and C7 corpectomy, autolog bone graft replacement of the two vertebral bodies, anterior plate fixation and posterior instrumentation with screws and rods). Postoperatively the patient had no significant pain, no neurological deficit and he was able to manage independent life himself. During the immediate follow-up CT of the neck showed the satisfactory position of the bone graft and the metal implantations. The 6 months follow-up CT revealed the anterior migration of the two screws from the Th1 vertebral body and 2 mm ventral elevation of the caudal end of the plate from the anterior surface of the Th1 vertebral body. The 1-year follow-up could not be performed because the patient died due to cardio-pulmonary insufficiency. This is the second Hungarian report of a chronic dialysis related severe spondylarthropathy which may cause pathologic fractures of the vertebral bodies. The typical radiological and histological findings are discussed. This disease affect patients’ quality of life and the conservative treatment alone seems to be ineffective in most cases. Based on the literature and personal experiences, the authors suggest 360-degree fixation of the spine to provide sufficient stability for the vertebrae of ”bad bone quality”, and early mobilisation of the patient can be achieved.

Összefoglaló. Az akut myeloid leukaemia ellátása primeren hematológiai feladat, relapsusa során is ritka a sebészeti szövődmény. Esettanulmányunkban egy 46 éves férfi beteget mutatunk be, akinél rutinvérvétel során felfedezett akut promyelocytás leukaemia miatt történt hematológiai kezelés. Alapbetegsége komplett molekuláris remisszióba jutott, ezt követően fenntartó kezeléseket kapott, de betegsége kiújult. Reindukciós kezelést követően fehérvérsejtszáma normalizálódott, de nem jutott molekuláris remisszióba. További kezelés után a férfi testvérétől gyűjtött őssejttel allogén csontvelő-transzplantáció történt. Közel egy éve molekuláris remisszióban volt a beteg, amikor jelentkeztek jobb alhasi panaszai neutropenia, a C-reaktív protein magas értéke és pozitív hasi ultrahang mellett. Akutan laparoszkópos módszerrel távolítottuk el a láthatóan gyulladt féregnyúlványt. A hisztológiai feldolgozás során az akut promyelocytás leukaemia manifesztációja igazolódott. Tekintettel arra, hogy közben csontvelői relapsus is kialakult, újabb reindukciós kezelés, és másik testvérétől vett őssejtekkel ismételten allogén csontvelő-transzplantáció történt. Az általunk ismert irodalomban appendicitis képében jelentkező akut promyelocytás leukaemia relapsusáról nem találtunk közleményt, ezért tartottuk esetünket publikációra érdemesnek. Orv Hetil. 2020; 161(51): 2171–2174. Summary. The treatment of acute myeloid leukemia is primarily hematological. Surgical complications are rare even during relapse. We present the case of a 46-year-old man who was diagnosed with acute promyelocytic leukemia after a routine blood test. Following hematological treatment, molecular remission was achieved, and he was on maintenance therapy, when his leukemia relapsed. He received re-induction treatment and his white blood cell count normalised, however, molecular remission was not reached. After further treatment, allogeneic bone marrow transplantation was performed with stem cells collected from his brother. He developed right lower quadrant abdominal pain with neutropenia, elevated C-reactive protein and abdominal ultrasound report showed thickened appendix after nearly one year of molecular remission. We performed en emergency laparoscopic appendicectomy to remove the inflamed appendix. Histology confirmed acute promyelocytic leukemia manifestation in the appendix. Given the repeated bone marrow relapse, he received re-induction treatment and allogeneic bone marrow transplantation again with stem cells collected from his another brother. To the best of our knowledge, no previous report in the literature can be found of appendicitis mimicking relapse of acute promyelocytic leukemia, thus, this case report holds merit for publication. Orv Hetil. 2020; 161(51): 2171–2174.

Összefoglaló. Az acut myeloid leukemia a csontvelői eredetű hemopoetikus őssejtek malignus klonális betegsége. A betegség lefolyása, különösen a nagy dózisú kemoterápiára és allogén csontvelő-transzplantációra alkalmatlan betegek számára, a legutóbbi időkig szinte kivétel nélkül fatális volt. Az idős és/vagy leromlott állapotú acut myeloid leukemiás betegek kezelésében óriási áttörést hozott a Bcl2- (B-cell lymphoma 2) inhibitor – venetoclax és a hypomethyláló azacitidin kombinációja. Ötvenéves nőbetegünkön a normál karyotípusú, nucleophosmin 1 mutációval járó acut myeloid leukemia diagnózisa előtt szeptikus állapotot okozó végbéltályog miatt tehermentesítő colostomaképzést végeztünk. A széles spektrumú kombinált antibiotikus kezelés mellett is súlyosan elesett, szeptikus állapotú betegen a nagy dózisú, kuratív indukciós kezelést nem tartottuk kivitelezhetőnek, ezért venetoclax–azacitidin kombinációt alkalmaztunk. A beteg az első ciklus alatt az antibiotikus, antimycoticus kezelés mellett masszív hemoszupportációra szorult. A második ciklus alatt lényeges szövődményt már nem észleltünk, a ciklus végén elvégzett csontvelővizsgálat pedig komplett hematológiai remissziót igazolt. A páciens önellátóvá vált, a végbéltályog jelentősen javult. Az új kezelési lehetőség mellett elért remisszió, a beteg javuló általános állapota korábban ismeretlen problémát vet fel. Egy 50 éves beteg jó prognózisú eltéréssel járó acut myeloid leukemiájának ellátása a nagy dózisú kemoterápiával elért első remisszióban nem igényel allogén csontvelő-transzplantációt. „Nem kuratív” kezeléssel elért remisszióban hogyan folytassuk a kezelést? Summary. Acute myeloid leukemia is a clonal malignancy of bone marrow haemopoietic stem cells. Until recently the outcome of the disease has been almost without exemption fatal, especially in cases ineligible for high dose induction chemotherapy and bone marrow transplantation. The Bcl-2 inhibitor venetoclax-azacitidine combination is a breakthrough therapy for the elderly and frail AML patient. Prior to the diagnosis of normal karyotype NPM 1 mutant AML of our 50-year-old female patient a colostomy had been performed due to anal abscess leading to sepsis. The septic patient being extremely frail in spite of wide spectrum antibiotic treatment was deemed ineligible for high dose curative induction treatment, hence venetoclax-azacitidine combined treatment was used. During cycle 1 the patient needed massive transfusion support beside concomitant antibiotic and antimycotic medication. On course of cycle 2 no major complications were detected, BM evaluation at the end of cycle confirmed complete haemotologic remission. The abscess significantly improved, the patient’s self-care capability was restored. The remission induced by this new option and the improvement of the performance status of the patient raised hitherto unasked questions. In case of a patient aged 50, a favourable prognosis AML in remission following high dose induction does not mandate HSCT as a next step. How shall we follow the course of treatment in remission elicited by a ‘non-curative’ option?

Head and neck squamous cell carcinoma (HNSCC) is the sixth most common cancer worldwide. To improve pre- and post-operative diagnosis and prognosis novel molecular markers are desirable. Here we used MALDI imaging mass spectrometry (IMS) and immunohistochemistry (IHC) to seek tumor specific expression of proteins and lipids in HNSCC samples. Among low molecular weight proteins visualized, S100A8 and S100A9 were found to be expressed in the regions of tumor tissue but not in the surrounding healthy stroma of a post-operative microdissected tissue. Marker potential of S100A8 and S100A9 was confirmed by immunohistochemistry of paraffin-embedded pathological samples. Imaging lipids showed a remarkable depletion of lysophosphatidylcholine species LPC[16:0], LPC[18:2] and, in parallel, accumulation of major glycerophospholipid species PE-P[36:4], PC[32:1], PC[34:1] in neoplastic areas. This was confirmed by shotgun lipidomics of dissected healthy and tumor tissue sections. A combination of the negative (LPC[16:0]) and positive (PC[32:1], PC[34:1]) markers was also applicable to uncover tumorous character of a pre-operative biopsy. Furthermore, marker potential of lysophospholipids was supported by elevated expression levels of the lysophospholipid degrading enzyme lysophospholipase A1 (LYPLA1) in the tumor regions of paraffin-embedded HNSCC samples. Finally, experimental evidence of 3D cell spheroid tests showed that LPC[16:0] facilitates HNSCC invasion, implying that HNSCC progression in vivo may be dependent on lysophospholipid supply. Altogether, a series of novel proteins and lipid species were identified by IMS and IHC screening, which may serve as potential molecular markers for tumor diagnosis, prognosis, and may pave the way to better understand HNSCC pathophyisiology.

Absztrakt: A tirotropintermelő hypophysistumor a hyperthyreosis ritka oka, a hypophysisadenomák 1 százalékában fordul elő. Az utóbbi években a felismert esetek száma nőtt az ultraszenzitív immunometriás esszék elterjedésének köszönhetően. A klinikai gyakorlatban a normál- vagy emelkedett tirotropinérték mellett észlelt emelkedett perifériás pajzsmirigyhormonszintek esetében kell felvetni a tirotropintermelő tumor lehetőségét. A szerzők egy középkorú nőbeteg esetét ismertetik, akinél a centrális hyperthyreosis hátterében tirotropintermelő hypophysis-macroadenoma igazolódott. Átmeneti thyreostaticus kezelést követően a tumor radikális idegsebészeti eltávolítása megtörtént. A műtétet követően a beteg hyperthyreosisa megszűnt, a residualis hypophysisfunkció megtartott maradt. Gondozása során recidívára utaló eltérést nem észleltünk. Esetismertetésünkben a kórkép felismerése kapcsán felmerülő differenciáldiagnosztikai nehézségeket is taglaljuk. Orv Hetil. 2020; 161(12): 474–478.

Congenital glioblastoma (cGBM) is a brain tumor very rarely observed in newborns and young infants, and differs in several respects from glioblastoma (GBM) of childhood and adulthood. Our aim was the presentation of a cGBM case with 14 days of postnatal survival at the Pediatric Oncology Center of the Markusovszky University Teaching Hospital in 2004. We investigated formalin-fixed, paraffin-embedded autoptic tumor samples of the newborn by immunohistochemical and molecular genetic (FISH and pyrosequencing) methods. We found polysomy of chromosome 2 and 5' deletion of the ALK gene in the glioma cells by ALK FISH. This result indicates the importance of molecular analyses in the diagnostic evaluation of cGBM, and raises the possibility of a personalized, targeted therapy (crizotinib, alectinib).

BACKGROUND: Recent studies proved that metabolic changes in malignant disorders have an impact on protein glycosylation, however, only a few attempts have been made so far to use O-GlcNAc analysis as a prognostic tool. Glucose metabolism is reported to be altered in hematological malignancies thus, we hypothesized that monitoring intracellular O-GlcNAc levels in Rai stage 0-I (Binet A) CLL patients could give deeper insights regarding subtle metabolic changes of progression which are not completely detected by the routine follow-up procedures. OBJECTIVE: In this proof of concept study we established a flow cytometric detection method for the assessment of O-GlcNAcylation as a possible prognostic marker in CLL malignancy which was supported by fluorescence microscopy. METHODS: Healthy volunteers and CLL patients were recruited for this study. Lymphocytes were isolated, fixed and permeabilised by various methods to find the optimal experimental condition for O-GlcNAc detection by flow cytometry. O-GlcNAc levels were measured and compared to lymphocyte count and various blood parameters including plasma glucose level. RESULTS: The protocol we developed includes red blood cell lysis, formalin fixation, 0.1% Tween 20 permeabilisation and employs standardized cell number per sample and unstained controls. We have found significant correlation between O-GlcNAc levels and WBC (R2= 0.8535, p< 0.0029) and lymphocyte count (R2= 0.9225, p< 0.0006) in CLL patients. Interestingly, there was no such correlation in healthy individuals (R2= 0.05664 for O-GlcNAc vs WBC and R2= 0.04379 for O-GlcNAc vs lymphocytes). CONCLUSION: Analyzing O-GlcNAc changes in malignant disorders, specifically in malignant hematologic diseases such as CLL, could be a useful tool to monitor the progression of the disease.

Absztrakt: A Gaucher-kor a leggyakrabban előfordulo lizoszomalis tarolasi betegseg, amiert a β-glucocerebrosidase enzim elegtelen műkodese a felelős. A betegseg ritka, incidenciaja 1/60 000, az ask...

A full-term male neonate presented with a left sided cervical lump at the level of the thyroid gland. Magnetic resonance imaging (MRI) showed a benign heterogeneous solid mass with lobulated margins. The tumor underwent complete excision. Histology revealed the diagnosis of chondromesenchymal hamartoma in ectopic thyroid tissue.

Abstract: Introduction: Plasma cell myeloma is a hematological malignancy with heterogeneous genomic landscape and diverse clinical course. Recurrent chromosomal and subchromosomal aberrations commonly occur in this entity and are associated with the pathogenesis and progression of the disease. The identification of these alterations aids genetic characterization, classification and prognostication of patients. Aim: Molecular cytogenetic investigations of plasma cell myeloma patients treated at the University of Pécs Clinical Center and János Balassa County Hospital of Tolna County, Szekszárd, between 2005 and 2018 were evaluated in our study. Method: 231 patients were screened for genetic aberrations using fluorescence in situ hybridization. Translocations involving the immunoglobulin heavy chain gene, losses of 1p and 17p chromosome arms, gains of 1q chromosome arm and unbalanced aberrations of chromosome 13 were investigated. Losses and gains of 1p, 1q, 5q, 12p, 13q, 16q and 17p chromosome arms were analyzed using multiplex ligation-dependent probe amplification in 42 patients. During the investigated period, 116 bone marrow karyotyping was also performed. Results: In total, 233 genetic aberrations were identified using our targeted approaches; the frequency of specific aberrations correlated with data of the recent literature. Concordance of results gained by fluorescence in situ hybridization and multiplex ligation-dependent probe amplification was 96.2% by analyzing the same chromosome arms. The latter technique revealed 21 additional genetic aberrations in 16/42 patient samples (38%) as compared to fluorescence in situ hybridization. Conclusions: Our results suggest that the combined application of the two molecular cytogenetic methods may facilitate a more detailed characterization of genetic aberrations of plasma cell myeloma patients in Hungary. Orv Hetil. 2019; 160(24): 944–951.

Abstract: Introduction: Prognostic impact of the detection of minimal residual disease (MRD) in multiple myeloma (MM) has been confirmed in numerous studies. Aim: Retrospective examination of our patient database (107 newly diagnosed multiple myelomas between 2007 and 2017). Flow cytometry (FCM) was performed as MRD assessment. Method: MRD assessment was performed in 56 patients (median age: 68 years), after induction treatment of multiple myeloma. The treatment contained bortezomib in 91%, autologous haematopoetic stem cell transplantation (ASCT) was perfomed in 50%. MRD detection was performed on bone marrow samples, predominantly in our hospital (BD FACScan, 3 colour, panel: CD38, CD138, CD19, CD45, CD56, CD28, CD117, cyKappa, cyLambda, 100 000 events). Statistical analysis: SPSS 13.0. Results: The progression-free survival (PFS) and the overall survival (OS) were significantly longer in MRD negative (n = 22) patients (PFS: 54 months, OS: 79% after 5 years) than MRD positive patients (n = 34, PFS: 22 months, OS 21% after 5 years, p = 0.001). Patients achieving complete response (CR) (n = 29) have different PFS (MRD negative CR: 60 months, MRD positive CR: 21 months, p

Glioblastoma (GBM) is the most aggressive tumor of the central nervous system (CNS). The standard of care improves the overall survival of patients only by a few months. Explorations of new therapeutic targets related to molecular properties of the tumor are under way. Even though neurotransmitters and their receptors normally function as mediators of interneuronal communication, growing data suggest that these molecules are also involved in modulating the development and growth of GBM by acting on neuronal and glioblastoma stem cells. In our previous DNA CpG methylation studies, gene ontology analyses revealed the involvement of the monoamine pathway in sequential GBM. In this follow-up study, we quantitated the expression levels of four selected catecholamine pathway markers (alpha 1D adrenergic receptor—ADRA1D, adrenergic beta receptor kinase 1 or G protein-coupled receptor kinase 2—ADRBK1/GRK2, dopamine receptor D2—DRD2, and synaptic vesicle monoamine transporter—SLC18A2) by immunohistochemistry, and compared the histological scores with the methylation levels within the promoters + genes of these markers in 21 pairs of sequential GBM and in controls. Subsequently, we also determined the promoter and gene methylation levels of the same markers in an independent database cohort of sequential GBM pairs. These analyses revealed partial inverse correlations between the catecholamine protein expression and promoter + gene methylation levels, when the tumor and control samples were compared. However, we found no differences in the promoter + gene methylation levels of these markers in either our own or in the database primary–recurrent GBM pairs, despite the higher protein expression of all markers in the primary samples. This observation suggests that regulation of catecholamine expression is only partially related to CpG methylation within the promoter + gene regions, and additional mechanisms may also influence the expression of these markers in progressive GBM. These analyses underscore the involvement of certain catecholamine pathway markers in GBM development and suggest that these molecules mediating or modulating tumor growth merit further exploration.

Central nervous system (CNS) lymphoma is a rare and aggressive non-Hodgkin lymphoma that might arise in the CNS (primary CNS lymphoma, PCNSL) or disseminates from a systemic lymphoma to the CNS (secondary CNS lymphoma, SCNSL). Dysregulated expression of microRNAs (miRNAs) is associated with various pathological processes and miRNA expression patterns may have diagnostic, prognostic and therapeutic implications. However, miRNA expression is understudied in CNS lymphomas. Here, we performed expression analysis of 798 miRNAs in 73 CNS lymphoma samples using the NanoString platform, followed by a detailed statistical analysis to identify potential novel biomarkers characterizing subgroups and to examine differences based on their primary and secondary nature, molecular subtype, mutational patterns and survival. We describe the general expression patterns of miRNAs across CNS lymphoma samples and identified 31 differentially expressed miRNAs between primary and secondary groups. Additionally, we identified 7 more miRNAs associated with a molecular subtype and 25 associated with mutation status. Using unsupervised clustering methods, we defined a small but distinct primary CNS lymphoma subgroup, with characteristically different expression patterns compared to the rest of the cases. Finally, we identified differentially regulated pathways in the above comparisons and assessed the utility of miRNA expression patterns in predicting survival. Our study identifies a novel CNS lymphoma subgroup defined by distinct miRNAs, proves the importance of specific miRNAs and pathways in their pathogenesis, and provides the basis for future research.

The Bruton's tyrosine kinase (BTK) inhibitor ibrutinib has revolutionised the therapeutic landscape of chronic lymphocytic leukaemia (CLL). Acquired mutations emerging at position C481 in the BTK tyrosine kinase domain are the predominant genetic alterations associated with secondary ibrutinib resistance. To assess the correlation between disease progression, and the emergence and temporal dynamics of the most common resistance mutation BTK

The Bruton's tyrosine kinase (BTK) inhibitor ibrutinib has revolutionised the therapeutic landscape of chronic lymphocytic leukaemia (CLL). Acquired mutations emerging at position C481 in the BTK tyrosine kinase domain are the predominant genetic alterations associated with secondary ibrutinib resistance. To assess the correlation between disease progression, and the emergence and temporal dynamics of the most common resistance mutation BTKC481S , sensitive (10-4 ) time-resolved screening was performed in 83 relapsed/refractory CLL patients during single-agent ibrutinib treatment. With a median follow-up time of 40 months, BTKC481S was detected in 48·2% (40/83) of the patients, with 80·0% (32/40) of them showing disease progression during the examined period. In these 32 cases, representing 72·7% (32/44) of all patients experiencing relapse, emergence of the BTKC481S mutation preceded the symptoms of clinical relapse with a median of nine months. Subsequent Bcl-2 inhibition therapy applied in 28/32 patients harbouring BTKC481S and progressing on ibrutinib conferred clinical and molecular remission across the patients. Our study demonstrates the clinical value of sensitive BTKC481S monitoring with the largest longitudinally analysed real-world patient cohort reported to date and validates the feasibility of an early prediction of relapse in the majority of ibrutinib-treated relapsed/refractory CLL patients experiencing disease progression.

Background Wnt signaling plays important roles in tumorigenesis, invasiveness and therapeutic resistance of glioblastoma (GBM). Methods We simultaneously investigated six Wnt pathway markers (Wnt5a, Fzd-2, beta-catenin, Wnt3a, Wnt7b, Fzd-10) at epigenetic and protein levels in 21 sequential formalin-fixed paraffin-embedded GBM pairs and controls. Results Expression levels of Wnt5a, beta-catenin and Wnt3a proteins either moderately or significantly increased, while those of Fzd-2, Wnt7b and Fzd-10 decreased in the primary (GBM-P) and recurrent (GBM-R) tumors compared to the controls. Methylation levels within promoters and genes showed corresponding decreases for Wnt5a, beta-catenin and Wnt3a in tumors vs. controls, while that of Fzd-10 was uniformly high. Comparing the GBM-P and GBM-R pairs, proteins of Fzd-2, beta-catenin and Wnt3a were either moderately or significantly up-, while that of Wnt7b was downregulated in GBM-R, but these patterns were not accompanied by inverse methylation patterns in the corresponding promoters and genes over time. No methylation differences were noted within promoters and genes of the same markers in 112 pairs of primary and recurrent GBMs in a database, suggesting that the observed changes in protein expression levels may not be explained by CpG methylation status alone. The promoter and gene methylation rate was the highest for Fzd-10 in the database cohort too, supporting the noted low Fzd-10 protein expression. Discussion These analyses underscore the relevance of Wnt pathway molecules in the context of their methylation profiles in the development and evolution of GBM, and suggest that Wnt pathway regulation as a potential treatment target merits further studies.

Purpose Glioblastoma is the most aggressive form of brain tumors. A better understanding of the molecular mechanisms leading to its evolution is essential for the development of treatments more effective than the available modalities. Here, we aim to identify molecular drivers of glioblastoma development and recurrence by analyzing DNA CpG methylation patterns in sequential samples. Methods DNA was isolated from 22 pairs of primary and recurrent formalin-fixed, paraffin-embedded glioblastoma specimens, and subjected to reduced representation bisulfite sequencing. Bioinformatic analyses were conducted to identify differentially methylated sites and pathways, and biostatistics was used to test correlations among clinical and pathological parameters. Results Differentially methylated pathways likely involved in primary tumor development included those of neuronal differentiation, myelination, metabolic processes, synapse organization and endothelial cell proliferation, while pathways differentially active during glioblastoma recurrence involved those associated with cell processes and differentiation, immune response, Wnt regulation and catecholamine secretion and transport. Conclusion DNA CpG methylation analyses in sequential clinical specimens revealed hypomethylation in certain pathways such as neuronal tissue development and angiogenesis likely involved in early tumor development and growth, while suggested altered regulation in catecholamine secretion and transport, Wnt expression and immune response contributing to glioblastoma recurrence. These pathways merit further investigations and may represent novel therapeutic targets.

Acute lymphoblastic leukemia is the most common pediatric cancer characterized by a heterogeneous genomic landscape with copy number aberrations occurring at various stages of pathogenesis, disease progression, and treatment resistance. In this study, disease-relevant copy number aberrations were profiled in bone marrow samples of 91 children with B- or T-cell precursor acute lymphoblastic leukemia using digital multiplex ligation-dependent probe amplification (digitalMLPA(TM)). Whole chromosome gains and losses, subchromosomal copy number aberrations, as well as unbalanced alterations conferring intrachromosomal gene fusions were simultaneously identified with results available within 36 hours. Aberrations were observed in 96% of diagnostic patient samples, and increased numbers of copy number aberrations were detected at the time of relapse as compared with diagnosis. Comparative scrutiny of 24 matching diagnostic and relapse samples from 11 patients revealed three different patterns of clonal relationships with (i) one patient displaying identical copy number aberration profiles at diagnosis and relapse, (ii) six patients showing clonal evolution with all lesions detected at diagnosis being present at relapse, and (iii) four patients displaying conserved as well as lost or gained copy number aberrations at the time of relapse, suggestive of the presence of a common ancestral cell compartment giving rise to clinically manifest leukemia at different time points during the disease course. A newly introduced risk classifier combining cytogenetic data with digitalMLPA(TM)-based copy number aberration profiles allowed for the determination of four genetic subgroups of B-cell precursor acute lymphoblastic leukemia with distinct event-free survival rates. DigitalMLPA(TM) provides fast, robust, and highly optimized copy number aberration profiling for the genomic characterization of acute lymphoblastic leukemia samples, facilitates the decipherment of the clonal origin of relapse and provides highly relevant information for clinical prognosis assessment.

Objectives Targeted therapies in the management of patients with lung cancer provide significantly better outcome compared to chemotherapy. Detection of the anaplastic lymphoma kinase (ALK) gene rearrangement has great predictive value for treatment with small molecule tyrosine kinase inhibitor (crizotinib and alectinib commonly). Fluorescent in situ hybridisation (FISH) assay is a basic diagnostic test designed for detecting ALK gene rearrangements. Although being considered as gold standard method by IASLC's guideline, it is often regarded as difficult and error prone. Our aim was to examine a unique atypical ALK FISH pattern, revealed during a systematic large-scale monitoring, which carries the great risk of misinterpretation, hence may result in loss of patients eligible for targeted therapy. Materials and Methods Tissue and cytology samples from nearly one thousand patients with advanced stage non-small cell lung cancer (NSCLC, n = 996) were routinely examined by ALK FISH and immunohistochemistry (Ventana ALK-D5F3-CDx assay). Anchored Multiplex PCR based Next Generation Sequencing (AMP-NGS) was used to detect fusion gene transcripts in ambiguous cases. Results Fifty-nine (5,9%) of the cases were positive with ALK FISH test. Three cases showed atypical pattern with a significantly reduced sized red (3′) signal and complete loss of green signals. Digital signal measurement confirmed this finding, showing consistent attenuation of 3′ signals throughout the tumours. In all three cases AMP-NGS and ALK IHC verified the presence of a fusion gene and expressed oncoprotein, respectively. Conclusion Approximately 5% of the 59 ALK positive cases exhibited atypical attenuated isolated 3′ signal pattern. The immunohistochemistry and AMP-NGS examinations helped to clarify the presence of oncoprotein and the fusion gene, respectively. Our results emphasize the importance of extensive exploration of the genetic background of any unexpected FISH finding to avoid false diagnosis. This enables clinicians to indicate the adequate therapy with higher efficiency for patients suffering from NSCLC.

Thyrotropin-secreting pituitary tumors are rare causes of hyperthyroidism and account for less than 1% of all pituitary adenomas. The number of reported cases increased over the last few years as a consequence of the routine use of ultrasensitive immunometric assays for measuring thyrotropin levels. In the clinical practice, thyrotropin secreting adenomas must be considered in case of inappropriately normal to elevated thyrotropin in the presence of elevated free serum thyroid hormone levels. The authors present the case history of a middle aged female patient, who suffered from hyperthyreodism caused by a thyrotropin-secreting pituitary macroadenoma. After transient thyreostatic treatment, radical neurosurgical removal of the tumor was performed. The pituitary surgery was effective in restoring the patient's euthyreodism. The postoperative pituitary function remained intact. During follow-up, the recurrence of the disease was not detected. In our case report, the difficulties in the differential diagnoses are also discussed. Orv Hetil. 2020; 161(12): 474-478.Absztrakt: A tirotropintermelő hypophysistumor a hyperthyreosis ritka oka, a hypophysisadenomák 1 százalékában fordul elő. Az utóbbi években a felismert esetek száma nőtt az ultraszenzitív immunometriás esszék elterjedésének köszönhetően. A klinikai gyakorlatban a normál- vagy emelkedett tirotropinérték mellett észlelt emelkedett perifériás pajzsmirigyhormonszintek esetében kell felvetni a tirotropintermelő tumor lehetőségét. A szerzők egy középkorú nőbeteg esetét ismertetik, akinél a centrális hyperthyreosis hátterében tirotropintermelő hypophysis-macroadenoma igazolódott. Átmeneti thyreostaticus kezelést követően a tumor radikális idegsebészeti eltávolítása megtörtént. A műtétet követően a beteg hyperthyreosisa megszűnt, a residualis hypophysisfunkció megtartott maradt. Gondozása során recidívára utaló eltérést nem észleltünk. Esetismertetésünkben a kórkép felismerése kapcsán felmerülő differenciáldiagnosztikai nehézségeket is taglaljuk. Orv Hetil. 2020; 161(12): 474–478.

Absztrakt: Bevezetes: A primer csontlimfomak rendkivul ritka entitasok, melyek patogeneziset illetően az alacsony esetszam miatt meg mindig keves informacio all rendelkezesunkre. A primer csontlimf...

Absztrakt: Az X kromoszomahoz kotott limfoproliferativ betegseg (XLP) ritka immundeficienciaval jaro szindroma, melyet az SH2D1A gen mutacioja okoz. A betegseget az Epstein–Barr-virusfertőzes aktiv...

EML4-ALK gene fusion (inv2(p21p23)) of non-small cell lung cancer (NSCLC) predisposes to tyrosine kinase inhibitor treatment. One of the gold standard diagnostics is the dual color (DC) break-apart (BA) FISH technique, however, the unusual closeness of the involved genes has been suggested to raise likelihood of random co-localization (RCL) of signals. Although this is suspected to decrease sensitivity (often to as low as 40-70%), the exact level and effect of RCL has not been revealed thus far. Signal distances were analyzed to the 0.1 µm precision in more than 25,000 nuclei, via automated high content-image cytometry. Negative and positive controls were created using conventional DC BA-, and inv2(p21p23) mimicking probe-sets, respectively. Average distance between red and green signals was 9.72 pixels (px) (±5.14px) and 3.28px (±2.44px), in positives and negatives, respectively; overlap in distribution being 41%. Specificity and sensitivity of correctly determining ALK status was 97% and 29%, respectively. When investigating inv2(p21p23) with DC BA FISH, specificity is high, but seven out of ten aberrant nuclei are inevitably falsely classified as negative, due to the extreme level of RCL. Together with genetic heterogeneity and dilution effect of non-tumor cells in NSCLC, this immense analytical false negativity is the primary cause behind the often described low diagnostic sensitivity. These results convincingly suggest that if FISH is to remain a gold standard for detecting the therapy relevant inv(2), either a modified evaluation protocol, or a more reliable probe-design should be considered than the current DC BA one. © 2018 International Society for Advancement of Cytometry.

Background Philadelphia negative myeloproliferative neoplasms (MPNs) are characterized by frequent mutations of driver genes including JAK2, CALR and MPL. While the influence of JAK2 V617F mutant allele burden on the clinical phenotype of MPN patients is well-described, the impact of CALR mutant allele burden on clinical features needs further investigation. Patients and methods Quantitative assessment of JAK2 and CALR mutations was performed on diagnostic DNA samples from 425 essential thrombocythemia (ET) and 227 primary myelofibrosis patients using real-time quantitative PCR and fragment length analysis. Characterization of CALR mutations and detection of MPL mutations were performed by Sanger sequencing. Results Twelve novel CALR mutations have been identified. ET patients with CALRmut load exceeding the median value exhibited lower hemoglobin values (12.0 vs. 13.6 g/dL), higher LDH levels (510 vs. 351 IU/L) and higher rate of myelofibrotic transformation (19% vs. 5%). The CALRmut load was higher among ET patients presenting with splenomegaly compared to those without splenomegaly (50.0% vs. 43.5%). Conclusion Our study confirms the clinical significance of driver mutational status and JAK2mut load in MPNs; in addition, unravels a novel clinical association between high CALRmut load and a more proliferative phenotype in ET.

Context Blastic plasmacytoid dendritic cell neoplasm (BPDCN) is a rare but highly aggressive hematologic malignancy derived from plasmacytoid dendritic cells. Currently, there is no consensus regarding the best therapeutic approach, and the diagnosis of BPDCN remains a challenge. Objective We aimed to collect and systematize the currently available evidence about the immunohistochemical (IHC) and flow cytometric (FCM) data in the diagnosis of BPDCN. Design A systematic search was carried out in MEDLINE (via PubMed), Embase, Cochrane Central Register of Controlled Trials, Scopus, Web of Science, China National Knowledge Infrastructure, and Latin American, and Caribbean Health Sciences Literature databases for studies published up to 20 April 2021. Patients or Other Participants We selected clinical studies reporting on CD4+/CD56+/CD123+ co-expression in BPDCN patients among other markers. Main Outcomes Measures Cell-surface marker expression assessed by either IHC or FCM was collected regarding 106 proteins. Results The systematic search and data collection are ongoing. Out of 3106 records collected, we identified 115 reported cases from 98 full-text articles. Conclusions Based on our preliminary findings, a confident immunophenotypic diagnosis can be made when CD4, CD56, CD123, TCL1, CD43, and CD45 are present and when other lineage-specific antigens are absent (CD19, CD20, CD3, MPO, and CD79a). This work is continuing in collecting all reported data about the diagnostic algorithm of this disorder, which could be used as a template for targeted therapy in BPDCN.

It has been suggested that hepatobiliary carcinomas are less frequent in Wilson’s disease (WD) than in liver diseases of other etiology. However, the protective role of copper against malignancies is debated. Only a few cases of cholangiocarcinoma (CCC) in WD have been published. Here we report on a case of a 47-year-old male H1069Q homozygous, Kayser-Fleischer ring positive WD patient with a low ceruloplasmin level who was followed up and treated with chelating agents throughout nine years. The patient presented with neurological symptoms and liver cirrhosis at diagnosis. Clinical symptoms regressed after the treatment initiation. Rapidly developed tumour metastases were found in the bones, lung and liver (without jaundice). Autopsy revealed cholangiocarcinoma as the primary tumour confirmed by strong CK7 positivity and glypican-3 negativity. The curiosity of the presented case is the very rapid development of CCC despite continuous chelating agent therapy.Abbreviations: CCC: cholangiocarcinoma; HCC: hepatocellular carcinoma; WD: Wilson disease.

Abstract: Introduction: In recent years much progress has been made in the therapy of chronic lymphocytic leukaemia, as the new innovative medicine proved to be effective in managing patients carrying TP53 abnormalities. To identify all these patients, it is essential to screen for both forms of TP53 defects, including both 17p deletions and TP53 mutations. Aim: The aim of this study was to determine the frequency of TP53 mutations and their association with 17p deletions in a large Hungarian cohort of 196 patients suffering from chronic lymphocytic leukaemia. Method: We performed mutation analysis of TP53 (exons 3–10) using Sanger sequencing. Results: TP53 mutations were present in 15.8% of patients, half of which were associated with 17p deletion. By analysing both forms, TP53 defect was identified in 25.4% of the patients. Conclusions: Our study demonstrates that by performing a TP53 mutation analysis, an additional 10% of high-risk patients can be detected. Orv. Hetil., 2017, 158(6), 220–228.

Despite the spectacular development of clinical immunotherapy (IT) in the last decade, the regular treatment approaches for the most common central nervous system (CNS) tumors, the malignant gliomas (MGs) has not changed yet. The most important pitfalls of the routine application of immunotherapy can be imputed to the special and originally immunosuppressed microenvironment and the extreme heterogeneity of MGs, however the defensive role of the blood-brain barrier, the general usage of steroids and the difficulties in the evaluation of brain images can also play a role in these types of difficulties. Additionally, in the case of MGs, well-accepted IT biomarker assays (PDL1 positivity, mismatch repair deficiencies, tumor mutation burden, etc.) generally reveal only minimal levels of immunogen activities. Nevertheless, there are some promising results with the utilization of checkpoint inhibitors and other IT modalities (such as virus-based therapies, tumor vaccines, adoptive T cell therapies) and with the combination of conventional oncotherapy methods in case of CNS malignancies, as well. In conclusion, although the CNS is not any more considered as an "immunological sanctuary" and despite some encouraging experimental and clinical results in CNS oncotherapy, the routine application of IT in case of MGs is still awaited.Az immunterápia elmúlt évtizedbeli látványos térnyerése ellenére a leggyakoribb primer idegrendszeri daganatok, a malignus gliómák esetében még nem változott meg a konvencionális onkoterápiás ellátás. Az immunterápia klinikai alkalmazásának eddigi kudarca talán leginkább az eredendõen immunszuppresszív mikrokörnyezeti tényezõknek és az extrém tumorheterogenitásnak tudható be, de ebben a bizonytalanságban szerepet játszhat a vér-agy gát védõ szerepe, agydaganatok esetében a szteroidok rutinszerû alkalmazása, és az idegrendszeri metszeti képalkotás sokszor igen bonyolult értékelhetõsége is. Mindezeken túlmenõen, a jelenleg ismert biomarkerek (PD-L1- és mismatch repair státusz, tumormutációs terheltség) gliómáknál szintén minimális immunogenitást jelzõ értékeket mutatnak. Pedig már bizonyos biztató klinikai eredmények ismertek malignus gliómák esetében is, mind az immunellenõrzõpont-gátlók, mind más immunterápiás modalitások (vírusalapú terápia, tumorvakcinák, adoptív T-sejt-terápia) esetében, illetve az immunterápia konvencionális kezelésekkel való kombinálásával. Összegezve elmondható, hogy bár az idegrendszer már nem tartható abszolút mértékben „immunológiai szentélynek”, és már vannak biztató kutatási és klinikai eredmények, a malignus gliómák immunkezelésének rutinszerû klinikai alkalmazása még várat magára.

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Introduction Two types of testicular teratomas are distinguished by the current WHO classification. Prepubertal-type teratomas are benign, while postpubertal-type teratomas are considered malignant with metastatic potential, and are associated with germ cell neoplasia in situ. Prepubertal-type cases have been reported in the adult testis potentially causing confusion and overtreatment. Demonstration of the absence of 12p abnormalities with fluorescence in situ hybridization may facilitate diagnosis. Recently, IMP3 has emerged as a potential marker of malignancy in this context. Aims The aim of this study was to assess histological characteristics, IMP3 expression and the presence of 12p abnormalities of pure testicular teratomas. Results Thirty-seven cases were studied, 7 patients were children and 30 were adults. Six out of 7 pediatric cases showed no 12p abnormality and were IMP3 positive. Seventy-four percent and 79% of adult cases showed 12p abnormalities and IMP3 expression, respectively. Negative cases were not associated with in situ neoplasia or metastasis, they were smaller (mean, 14 vs 39 mm), showed less histological diversity (2.4 vs 4.0 types of tissues on average) compared to positive cases. Conclusion Our study provides further evidence that prepubertal-type (type I) teratomas may appear in adult testes, thus teratomas in adults may be either benign (type I) or malignant (type II). IMP3 expression may aid the distinction between type I and type II teratomas of the postpubertal testis even when GCNIS and 12p status cannot be assessed.