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Introduction: The p.(Arg85Trp) variant-specific phenotype of hepatocyte nuclear factor 4 alpha shows a complex clinical picture affecting three different organ systems and their corresponding metabolisms. Little is known about the molecular mechanisms involved and their relationship with the diverse symptoms seen in the context of this specific variant. Here, we present data of a new patient that expand the clinical phenotype, suggesting possible disease mechanisms. Case Presentation: Clinical data were extracted from the patient’s charts. The liver, kidney, and muscle were analyzed with routine histology and electron microscopy. Mitochondrial function was assessed by respirometric analyses and enzymatic activity assays. Structure and sequence analyses of this specific variant were investigated by in silico analyses. Our patient showed the known features of the variant-specific phenotype, including macrosomia, congenital hyperinsulinism, transient hepatomegaly, and renal Fanconi syndrome. In addition to that, she showed liver cirrhosis, chronic kidney failure, and altered mitochondrial morphology and function. The clinical and biochemical phenotype had features of a new type of glycogen storage disease. Discussion: This case expands the p.(Arg85Trp) variant-specific phenotype. Possible pathomechanistic explanations for the documented multiorgan involvement and changes of symptoms and signs during development of this ultra-rare but instructive disorder are discussed.

Homozygous deletion (HD) of the CDKN2A/B locus has emerged as an unfavourable prognostic marker in diffuse gliomas, both IDH-mutant and IDH-wild-type. Testing for CDKN2A/B deletions can be performed by a variety of approaches, including copy number variation (CNV) analysis based on gene array analysis, next generation sequencing (NGS) or fluorescence in situ hybridisation (FISH), but questions remain regarding the accuracy of testing modalities. In this study, we assessed: (1) the utility of S-methyl-5'-thioadenosine phosphorylase (MTAP) and cellular tumour suppressor protein pl61NK4a (p16) immunostainings as surrogate markers for CDKN2A/B HD in gliomas, and (2) the prognostic value of MTAP, across different histological tumour grades and IDH mutation status. One hundred consecutive cases of diffuse and circumscribed gliomas (Cohort 1) were collected, in order to correlate MTAP and p16 expression with the CDKN2A/B status in the CNV plot of each tumour. IDH1 R132H, ATRX and MTAP immunohistochemistry was performed on next generation tissue microarrays (ngTMAs) of 251 diffuse gliomas (Cohort 2) for implementing survival analysis. Complete loss of MTAP and p16 by immunohistochemistry was 100% and 90% sensitive as well as 97% and 89% specific for CDKN2A/B HD, respectively, as identified on CNV plot. Only two cases (2/100) with MTAP and p16 loss of expression did not demonstrate CDKN2A/B HD in CNV plot; however, FISH analysis confirmed the HD for CDKN2A/B. Moreover, MTAP deficiency was associated with shortened survival in IDH-mutant astrocytomas (n=75; median survival 61 vs 137 months; p

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Metastases from primary prostate cancers to rare locations, such as the brain, are becoming more common due to longer life expectancy resulting from improved treatments. Epigenetic dysregulation is a feature of primary prostate cancer, and distinct DNA methylation profiles have been shown to be associated with the mutually exclusive SPOP-mutant or TMPRSS2-ERG fusion genetic backgrounds. Using a cohort of prostate cancer brain metastases (PCBM) from 42 patients, with matched primary tumors for 17 patients, we carried out a DNA methylation analysis to examine the epigenetic distinction between primary prostate cancer and PCBM, the association between epigenetic alterations and mutational background, and particular epigenetic alterations that may be associated with PCBM. Multiregion sampling of PCBM revealed epigenetic stability within metastases. Aberrant methylation in PCBM was associated with mutational background and PRC2 complex activity, an effect that is particularly pronounced in SPOP-mutant PCBM. While PCBM displayed a CpG island hypermethylator phenotype, hypomethylation at the promoters of genes involved in neuroactive ligand–receptor interaction and cell adhesion molecules such as GABRB3, CLDN8, and CLDN4 was also observed, suggesting that cells from primary tumors may require specific reprogramming to form brain metastasis. This study revealed the DNA methylation landscapes of PCBM and the potential mechanisms and effects of PCBM-associated aberrant DNA methylation.Significance:DNA methylation analysis reveals the molecular characteristics of PCBM and may serve as a starting point for efforts to identify and target susceptibilities of these rare metastases.

Metastases from primary prostate cancers to rare locations, such as the brain, are becoming more common due to longer life expectancy resulting from improved treatments. Epigenetic dysregulation is a feature of primary prostate cancer, and distinct DNA methylation profiles have been shown to be associated with the mutually exclusive SPOP-mutant or TMPRSS2-ERG fusion genetic backgrounds. Using a cohort of prostate cancer brain metastases (PCBM) from 42 patients, with matched primary tumors for 17 patients, we carried out a DNA methylation analysis to examine the epigenetic distinction between primary prostate cancer and PCBM, the association between epigenetic alterations and mutational background, and particular epigenetic alterations that may be associated with PCBM. Multiregion sampling of PCBM revealed epigenetic stability within metastases. Aberrant methylation in PCBM was associated with mutational background and PRC2 complex activity, an effect that is particularly pronounced in SPOP-mutant PCBM. While PCBM displayed a CpG island hypermethylator phenotype, hypomethylation at the promoters of genes involved in neuroactive ligand–receptor interaction and cell adhesion molecules such as GABRB3, CLDN8, and CLDN4 was also observed, suggesting that cells from primary tumors may require specific reprogramming to form brain metastasis. This study revealed the DNA methylation landscapes of PCBM and the potential mechanisms and effects of PCBM-associated aberrant DNA methylation. Significance: DNA methylation analysis reveals the molecular characteristics of PCBM and may serve as a starting point for efforts to identify and target susceptibilities of these rare metastases.

Background: The novel beta-coronavirus, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), enters the human body via mucosal surfaces of the upper and/or lower respiratory tract. Viral entry into epithelial cells is mediated via angiotensin-converting enzyme 2 (ACE2) and auxiliary molecules, but the precise anatomic site of infection still remains unclear. Methods: Here, we systematically investigated the main SARS-CoV-2 receptor proteins ACE2 and transmembrane serine protease 2 (TMPRSS2), as well as 2 molecules potentially involved in viral entry, furin and CD147, in formalin-fixed, paraffin-embedded human tissues. Tissue microarrays incorporating a total of 879 tissue cores from conjunctival (n = 84), sinonasal (n = 95), and lung (bronchiolar/alveolar; n = 96) specimens were investigated for protein expression by immunohistochemistry. Results: ACE2 and TMPRSS2 were expressed in ciliated epithelial cells of the conjunctivae and sinonasal tissues, with highest expression levels observed in the apical cilia. In contrast, in the lung, the expression of those molecules in bronchiolar and alveolar epithelial cells was much rarer and only very focal when present. Furin and CD147 were more uniformly expressed in all tissues analyzed, including the lung. Interestingly, alveolar macrophages consistently expressed high levels of all 4 molecules investigated. Conclusions: Our study confirms and extends previous findings and contributes to a better understanding of potential SARS-CoV-2 infection sites along the human respiratory tract.

Synoptic reporting increases completeness and standardization of surgical pathology reports and thereby contributes to an increased quality of clinical cancer care. Nevertheless, its widespread practical implementation remains a challenge, which is in part related to the effort required for setup and maintenance of database structures. This prompted us to assess the effect of a simple template-based, database-free system for synoptic reporting on completeness of surgical pathology reports. For this purpose, we analyzed 200 synoptic reports (100 colon and 100 lung cancer resections each) for completeness as required by the pertinent College of American Pathologists (CAP) protocols and compared these to a control dataset of 200 narrative reports. Introduction of template-based synoptic reporting resulted in improved completeness (98% of mandatory data elements) as compared to narrative reports (77%). Narrative reports showed a high degree of completeness for data elements covered by previously existing dictation templates. In conclusion, template-based synoptic reporting without underlying database structure can be a useful transitory phase in the implementation of synoptic reporting. It can result in a similar degree of completeness as reported in the literature for database solutions and provides other benefits of synoptic reporting while facilitating its implementation.

Synoptic reporting in pathology as opposed to traditional narrative reporting is defined by a laboratory value-like format and use of standardized checklists. It contributes to completeness and comprehensibility of pathology reports and ultimately patient care. As of today, two major institutions publish synoptic reporting templates, the College of American Pathologists (CAP) and the International Collaboration for Cancer Reporting (ICCR). Synoptic protocols are available for all major cancer types and provide not only a standardized terminology and a checklist for completeness of reports, but also facilitate uniform utilization of diagnostic criteria. Additionally, both CAP and ICCR protocols are accompanied by detailed and up-to-date reference lists and thereby represent a valuable source of information even when synoptic reporting is not used. The benefits and challenges of implementation of synoptic reporting are discussed, in particular with regard to reporting in German.Synoptische oder strukturierte Berichte in der Medizin, speziell in der Pathologie, sind im Gegensatz zu traditionellen narrativen Berichten gekennzeichnet durch ein listen- bzw. laborwertartiges Format und die Verwendung standardisierter Checklisten. Sie tragen zur Vollständigkeit und Verständlichkeit der Berichte und damit letztlich zu einer verbesserten Patientenversorgung bei. Für die Verwendung in der Pathologie publizieren aktuell 2 größere Institutionen Vorlagen für synoptische Berichte, das College of American Pathologists (CAP) und die International Collaboration for Cancer Reporting (ICCR). Synoptische Berichtsvorlagen sind für alle häufigeren Tumortypen verfügbar. Sie bieten nicht nur eine standardisierte Nomenklatur und Checklisten für vollständige Berichte, sondern unterstützen auch eine einheitliche Anwendung diagnostischer Kriterien. Außerdem beinhalten die Protokolle sowohl von CAP als auch von ICCR umfangreiche und aktuelle Referenzen und sind damit eine wertvolle Quelle von Zusatzinformationen, selbst wenn kein synoptisches Berichtsformat verwendet wird. Nutzen und Herausforderungen bei der Implementation von synoptischen Berichten werden diskutiert, insbesondere im Hinblick auf die deutschsprachige Pathologie.

Dasatinib, a second-generation tyrosine kinase inhibitor (TKI), used as treatment for chronic myeloid leukemia, BCR::ABL1-positive (CML), is complicated by pleural or pericardial effusions in about one-third of patients. Besides, in exceptional instances, effusion-based neoplastic B-cell lymphoproliferations have been described. Here, we report an HHV8-negative, EBV-positive large B-cell lymphoma presenting as a pericardial effusion in a patient with CML treated with dasatinib for 23 months, without associated tumor mass or lymphadenopathies. Large tumor cells showed a B-cell phenotype (CD20+, CD79+), with evidence of EBV infection (EBER-ISH+), but HHV8 (LANA-1) negative. Monoclonal IG gene rearrangements were identified. BCL2, BCL6, and MYC genes were not rearranged. Despite the aggressive cytomorphology the patient was in complete remission after 4 cycles of R-CHOP after 8 months follow-up. Four other cases of large B-cell effusion-based lymphomas developed in the setting of dasatinib therapy for CML have been reported in the literature. The four cases were HHV8-negative and one case was EBV-positive. Three of the four patients experienced a benign clinical course, which is in contrast to HHV8-positive primary effusion lymphoma (PEL). The mechanisms of development of these effusion-based B-cell lymphoproliferations in patients receiving TKI are not completely elucidated. Acute or relapsing effusions during TKI treatment in the setting of CML should be cytologically examined to exclude clonal B-cell lymphoproliferations.

Glioblastomas with methylation of the promoter region of the O(6)-methylguanine-DNA methyltransferase (We included consecutive patients ≥18 years treated at our department between 11/2010 and 08/2018 for a glioblastoma,A total of 321 patients were included. Median overall survival was 12.6 months. Kaplan-Meier and adjusted Cox regression analysis showed better survival for the groups with 16-30%, 31-60%, and 61-100% methylation. In contrast, survival in the group with 1-15% methylation was similar to those with unmethylated promoter regions. A secondary analysis confirmed this threshold.Better survival is observed in patients with glioblastomas with ≥16% methylation of the

(1) Background: Superior cerebellar peduncle (SCP) lesions are sparsely reported in the literature. The surgical approaches to the cerebello-mesencephalic region remain challenging. In this article, we present the extreme lateral supracerebellar infratentorial (ELSI) approach to treat a large hemorrhagic pilocytic astrocytoma of the SCP. (2) Methods: An 11-year-old boy, known for neurofibromatosis Type I, presented to the emergency department of our institution with symptoms and signs of intracranial hypertension. The cerebral magnetic resonance imaging (MRI) revealed a large hemorrhagic lesion centered on the SCP provoking obstructive hydrocephalus. Following an emergency endoscopic third ventriculocisternostomy (ETV), he underwent a tumor resection via an endoscope-assisted ELSI approach. (3) Results: ELSI approach allows for a wide exposure with direct access to lesions of the SCP. The post-operative course was uneventful, and the patient was discharged home on post-operative day 5. Post-operative MRI revealed a near total resection with a small residual tumor within the mesencephalon. (4) Conclusion: ELSI approach offers an excellent exposure with the surgical angles necessary for median and paramedian lesions. The park-bench position with appropriate head flexion and rotation offers a gravity-assisted relaxation of the tentorial and petrosal cerebellar surfaces. The endoscope can be an adjunct to illuminate the blind areas of the surgical corridor for an improved tumor resection without significant cerebellar retraction.

Activating mutations affecting exon 3 of the β-Catenin (CTNNB1) gene result in constitutive activation of WNT signalling and are a diagnostic hallmark of several tumour entities including desmoid-type fibromatosis or define clinically relevant subtypes such as in endometrioid carcinoma. In a diagnostic setting, β-Catenin immunohistochemistry is widely used as a surrogate of CTNNB1 mutations, but is often difficult to assess in practice, given that the characteristic nuclear translocation may be focal or hard to distinguish from spillover of the normal membranous staining.We therefore assessed Lymphoid Enhancer-Binding Factor 1 (LEF1) immunohistochemistry, a nuclear marker of WNT activation as a potential surrogate of CTNNB1 mutations. Across a variety of entities characterised by CTNNB1 mutations as a putative driver we found diffuse and strong expression of LEF1 in 77% of cases. In a cohort of endometrial carcinomas (n=255) LEF1 was accurate to predict CTNNB1 mutations in 85% (pWe conclude that LEF1 immunostaining is a highly useful surrogate marker of CTNNB1 mutations in lesions which are driven by WNT signalling, favourably complementing β-Catenin immunohistochemistry and outperforming the latter as a single marker.

Oligodendrogliomas are defined at the molecular level by the presence of an IDH mutation and codeletion of chromosomal arms 1p and 19q. In the past, case reports and small studies described gliomas with sarcomatous features arising from oligodendrogliomas, so called oligosarcomas. Here, we report a series of 24 IDH-mutant oligosarcomas from 23 patients forming a distinct methylation class. The tumors were recurrences from prior oligodendrogliomas or developed de novo. Precursor tumors of 12 oligosarcomas were histologically and molecularly indistinguishable from conventional oligodendrogliomas. Oligosarcoma tumor cells were embedded in a dense network of reticulin fibers, frequently showing p53 accumulation, positivity for SMA and CALD1, loss of OLIG2 and gain of H3K27 trimethylation (H3K27me3) as compared to primary lesions. In 5 oligosarcomas no 1p/19q codeletion was detectable, although it was present in the primary lesions. Copy number neutral LOH was determined as underlying mechanism. Oligosarcomas harbored an increased chromosomal copy number variation load with frequent CDKN2A/B deletions. Proteomic profiling demonstrated oligosarcomas to be highly distinct from conventional CNS WHO grade 3 oligodendrogliomas with consistent evidence for a smooth muscle differentiation. Expression of several tumor suppressors was reduced with NF1 being lost frequently. In contrast, oncogenic YAP1 was aberrantly overexpressed in oligosarcomas. Panel sequencing revealed mutations in NF1 and TP53 along with IDH1/2 and TERT promoter mutations. Survival of patients was significantly poorer for oligosarcomas as first recurrence than for grade 3 oligodendrogliomas as first recurrence. These results establish oligosarcomas as a distinct group of IDH-mutant gliomas differing from conventional oligodendrogliomas on the histologic, epigenetic, proteomic, molecular and clinical level. The diagnosis can be based on the combined presence of (a) sarcomatous histology, (b) IDH-mutation and (c) TERT promoter mutation and/or 1p/19q codeletion, or, in unresolved cases, on its characteristic DNA methylation profile.

Glial cell activation is a hallmark of several neurodegenerative and neuroinflammatory diseases. During HIV infection, neuroinflammation is associated with cognitive impairment, even during sustained long-term suppressive antiretroviral therapy. However, the cellular subsets contributing to neuronal damage in the CNS during HIV infection remain unclear. Using post-mortem brain samples from eight HIV patients and eight non-neurological disease controls, we identify a subset of CNS phagocytes highly enriched in LGALS3, CTSB, GPNMB and HLA-DR, a signature identified in the context of ageing and neurodegeneration. In HIV patients, the presence of this phagocyte phenotype was associated with synaptic stripping, suggesting an involvement in the pathogenesis of HIV-associated neurocognitive disorder. Taken together, our findings elucidate some of the molecular signatures adopted by CNS phagocytes in HIV-positive patients and contribute to the understanding of how HIV might pave the way to other forms of cognitive decline in ageing HIV patient populations.

We suggest using the wide-field imaging Mueller polarimetry to contrast optically anisotropic fiber tracts of healthy brain white matter for the detection of brain tumor borders during neurosurgery. Our prior studies demonstrate that this polarimetric imaging modality detects correctly the in-plane orientation of brain white matter fiber tracts of a flat formalin-fixed thick brain specimen in reflection geometry [IEEE Trans. Med. Imaging39, 4376 (2020)10.1109/TMI.2020.3018439]. Here we present the results of

In der stufenweisen Abklarung von Raumforderungen des Pankreas nimmt die Zytologie eine zentrale Rolle ein. Die Burstenzytologie und ultraschallgesteuerte endoskopische Feinnadelpunktionen liefern Material fur eine Diagnosestellung vor einer chirurgischen oder konservativen Therapie. Das Diagnosesystem der Papanicolaou Society of Cytopathology bildet einen konzeptuellen Rahmen fur die Befundung dieser Proben. Eine besondere Herausforderung in der Zytologie des Pankreas sind zystische Lasionen, da diese oftmals rein zytomorphologisch nicht adaquat diagnostiziert werden konnen. Aus konzeptueller Sicht wegweisend ist die Einbindung nichtmorphologischer Methoden im Papanicolaou Society System: Laborchemische (CEA >192 ng/ml) und molekulare (KRAS- und/oder GNAS-Mutation) Befunde an der Zystenflussigkeit sind Bestandteil der formalen diagnostischen Kriterien neoplastischer Zysten. Die Berner Erfahrungen zeigen, dass ein solcher integrativer Ansatz zu einem erheblichen Gewinn an diagnostischer Aussagekraft fuhrt. In einer Analyse von 88 Fallen konnte in 84 Fallen (95 %) adaquate DNA aus der Zystenflussigkeit extrahiert werden. Pathogene Mutationen waren mittels Next Generation Sequencing in 48 Fallen (55 %) nachweisbar. Davon ware in 35 Fallen (77 %) die Diagnose einer neoplastischen Zyste rein morphologisch nicht moglich gewesen. Diese Erfahrungen eroffnen neue Perspektiven fur die die Einbindung von Zusatztechniken an Stellen, wo die reine zytomorphologische Diagnostik an ihre Grenzen stost. In diesem Sinne sollte die Liquid Biopsy nicht als Ersatz, sondern als Erweiterung der zytologischen Diagnostik gesehen werden, entsprechend dem Motto „Doing more with less“.

AIMS The DNA mismatch repair (MMR) system is critical for genomic stability correcting errors generated during DNA replication with germline pathogenic variants (PVs) causing Lynch syndrome. Although acquired MMR deficiency is associated with temozolomide resistance, the incidence in treatment naïve tumours is unknown and not routinely screened for. Here we present three MMR deficient gliomas and describe their comprehensive molecular profile. METHOD Thirteen gliomas; 9/34 that had undergone sequencing of 50 cancer-associated genes and found to have frameshift/multiple point PVs, indicative of defective MMR, and four MSI-high tumours, identified following microsatellite analysis for 1p/19q co-deletion as part of routine care, were selected for analysis. MMR expression was then assessed by immunohistochemistry. Seven tumours showed loss of one or more MMR proteins and were molecularly characterised using PCR-based Promega MSI analysis and NGS-based Illumina TruSight500 panel. RESULTS Pathogenic variants in MMR genes and hypermutated phenotypes (TMB > 10mutations/Mb) were identified in three cases. One individual harboured an additional POLE PV resulting in an ultra-hypermutated phenotype (>100mutations/Mb). Two MMR PVs were germline, leading to a new diagnosis of Lynch syndrome. Only one of three tumours was found to be MSI-high using the Promega MSI analysis whilst all three were identified as MSI-high using TruSight500 MSI calling algorithms. CONCLUSION We describe the comprehensive molecular profile of seven treatment naïve gliomas, three being MMR deficient. We show TruSight500 to be more sensitive and reliable for detecting MSI compared to Promega. Given the implications, this additional analysis should be considered as part of the molecular report on all gliomas.

The crucial problem of brain tumor surgery is the accurate detection the tumor border for safe and complete tumor resection. Whereas it is quite easy to identify brain tumor in preoperative magnetic resonance imaging, it is often difficult to differentiate solid tumor tissue from infiltrated white matter during surgery with conventional surgical intra-operative microscope. To address this problem we suggest exploring the optical anisotropy of healthy brain white matter which represents a highly ordered structure consisting of axons that are joined together in fiber tracts. Tumor cells grow chaotically and erase the optical anisotropy of healthy brain. Instead of detecting the tumor itself, we suggest to visualize healthy white matter by means of its fiber tracts by detecting the optical anisotropy of brain tissue. For this purpose we used a wide-field imaging Mueller polarimetric system operating in the visible wavelength range in backscattering configuration. The Mueller matrix images of the thick (~1cm) fixed human brain specimen and thick (~1cm) fresh veal brain specimen were measured at 633 nm in reflection. Lu Chipman decomposition was applied pixel-wise to the experimental Mueller matrices. The maps of azimuth of fast optical axis of linear birefringent medium showed a compelling correlation with the fiber tracts directions on histology image of thin whole mount silver-stained brain tissue section, that is gold standard for ex-vivo brain fiber tract visualization. Thus, label-free non-contact imaging Mueller polarimetry shows potential for the intra-operative visualization of brain white matter fiber tracts. Further studies are ongoing.

Parkinson’s disease is mainly characterized by a progressive loss of dopaminergic neurons in the substantia nigra pars compacta. Together with the small number, the high vulnerability of the dopaminergic neurons is a major pathogenic culprit of Parkinson’s disease. Our previous findings of a higher survival of dopaminergic neurons in the substantia nigra co-expressing Nogo-A in an animal model of Parkinson’s disease suggested that Nogo-A may be associated with dopaminergic neurons resilience against Parkinson’s disease neurodegeneration. In the present study, we have addressed the expression of Nogo-A in the dopaminergic neurons in the substantia nigra in postmortem specimens of diseased and non-diseased subjects of different ages. For this purpose, in a collaborative effort we developed a tissue micro array (TMA) that allows for simultaneous staining of many samples in a single run. Interestingly, and in contrast to the observations gathered during normal aging and in the animal model of Parkinson’s disease, increasing age was significantly associated with a lower co-expression of Nogo-A in nigral dopaminergic neurons of patients with Parkinson’s disease. In sum, while Nogo-A expression in dopaminergic neurons is higher with increasing age, the opposite is the case in Parkinson’s disease. These observations suggest that Nogo-A might play a substantial role in the vulnerability of dopaminergic neurons in Parkinson’s disease.