Paediatric multiple sclerosis and acute disseminated encephalomyelitis in Germany: results of a nationwide survey

The aim of this study was to evaluate the incidence of paediatric multiple sclerosis (MS) and acute disseminated encephalomyelitis (ADEM) in Germany. In a prospective nationwide survey carried out between 1997 and 1999, all registered new cases of paediatric MS and ADEM with an onset before the age of 16 years were evaluated using a standardised questionnaire. A total of 132 patients with suspected or definite MS and 28 patients with an assumed diagnosis of ADEM were reported. Among these, 82% of the MS patients were 10 years of age or older, as opposed to 18% in the ADEM-cohort. The female-to-male ratio was 1.2:1 in the MS-cohort and 0.8:1 in the ADEM-cohort. Manifestation was polysymptomatic in 67% of the MS patients compared to 86% of the ADEM patients. The most frequent primary symptoms in the MS-cohort were cerebellar (44%), sensory (39%) or visual (36%), followed by brainstem (30%), pyramidal (29%) and cerebromental (22%) complaints. Conclusion: The incidence of paediatric MS in Germany is more than fourfold higher than that of paediatric ADEM; in addition, it shows a strikingly different age-distribution. With an estimated minimum of 50 new cases per year, the incidence of paediatric MS in Germany is much more frequent than previously believed. This paper is dedicated to Prof. Dr. Helmut Bauer for his continuing encouragement and support of research in paediatric MS.     

Consensus Recommendations of the German Consortium for Hereditary Breast and Ovarian Cancer

BackgroundThe German Consortium for Hereditary Breast and Ovarian Cancer (GC-HBOC) has established a multigene panel (TruRisk®) for the analysis of risk genes for familial breast and ovarian cancer.SummaryAn interdisciplinary team of experts from the GC-HBOC has evaluated the available data on risk modification in the presence of pathogenic mutations in these genes based on a structured literature search and through a formal consensus process.Key MessagesThe goal of this work is to better assess individual disease risk and, on this basis, to derive clinical recommendations for patient counseling and care at the centers of the GC-HBOC from the initial consultation prior to genetic testing to the use of individual risk-adapted preventive/therapeutic measures.     

Update Breast Cancer 2022 Part 2 – Advanced Stage Breast Cancer

For patients with advanced breast cancer, several novel therapies have emerged in recent years, including CDK4/6 inhibitors, immune checkpoint inhibitors, PARP inhibitors, alpelisib, tucatinib and trastuzumab-deruxtecan, and sacituzumab-govitecan, which have transformed and expanded the therapeutic landscape for patients with advanced breast cancer. Some of these substances have now been approved for use in the early stages of the disease, or are expected to be approved in the near future, so the therapeutic landscape will change once again. Therefore, current scientific efforts are focused on the introduction of new substances and understanding their mechanisms of progression and efficacy. This review summarizes recent developments with reference to recent publications and conferences. Findings on the treatment of patients with HER2-positive breast cancer and brain metastases are presented, as are a number of studies looking at biomarkers in patients with HER2-negative, hormone receptor-positive breast cancer. In particular, the introduction of oral selective estrogen receptor degraders provides new opportunities to establish biomarker-based therapy. Molecular diagnostics is establishing itself as a diagnostic marker and parameter of progression.Key words: breast cancer, metastatic, biomarker, PD-L1, ADC     

Zebrafish model for allogeneic hematopoietic cell transplantation not requiring preconditioning

Recent work on vertebrate hematopoiesis has uncovered the presence of deeply rooted similarities between fish and mammals at molecular and cellular levels. Although small animal models such as zebrafish are ideally suited for genetic and chemical screens, the study of cellular aspects of hematopoietic development in lower vertebrates is severely hampered by the complex nature of their histocompatibility-determining genes. Hence, even when hosts are sublethally irradiated before hematopoietic cell transplantation, stable and long-term reconstitution by allogeneic stem cells often fails. Here, we describe the unexpected observation that transplantation and maintenance of allogeneic hematopoietic stem cells in zebrafish homozygous for the c-myb^t25127 allele, carrying a missense mutation (Ile181Asn) in the DNA binding domain can be achieved without prior conditioning. Using this model, we examined several critical parameters of zebrafish hematopoiesis in a near-physiological setting. Limiting dilution analysis suggests that the kidney marrow of adult zebrafish harbors about 10 transplantable hematopoietic stem cells; this tissue also contains thymus-settling precursors that colonize the thymic rudiment within days after transplantation and initiate robust T-cell development. We also demonstrate that c-myb mutants can be stably reconstituted with hematopoietic cells carrying specific genetic defects in lymphocyte development, exemplifying one of the many potential uses of this model in experimental hematology.     

Characterizing the “fungal shunt”: Parasitic fungi on diatoms affect carbon flow and bacterial communities in aquatic microbial food webs

Microbial interactions in aquatic environments profoundly affect global biogeochemical cycles, but the role of microparasites has been largely overlooked. Using a model pathosystem, we studied hitherto cryptic interactions between microparasitic fungi (chytrid Rhizophydiales), their diatom host Asterionella, and cell-associated and free-living bacteria. We analyzed the effect of fungal infections on microbial abundances, bacterial taxonomy, cell-to-cell carbon transfer, and cell-specific nitrate-based growth using microscopy (e.g., fluorescence in situ hybridization), 16S rRNA gene amplicon sequencing, and secondary ion mass spectrometry. Bacterial abundances were 2 to 4 times higher on individual fungal-infected diatoms compared to healthy diatoms, particularly involving Burkholderiales. Furthermore, taxonomic compositions of both diatom-associated and free-living bacteria were significantly different between noninfected and fungal-infected cocultures. The fungal microparasite, including diatom-associated sporangia and free-swimming zoospores, derived ∼100% of their carbon content from the diatom. By comparison, transfer efficiencies of photosynthetic carbon were lower to diatom-associated bacteria (67 to 98%), with a high cell-to-cell variability, and even lower to free-living bacteria (32%). Likewise, nitrate-based growth for the diatom and fungi was synchronized and faster than for diatom-associated and free-living bacteria. In a natural lacustrine system, where infection prevalence reached 54%, we calculated that 20% of the total diatom-derived photosynthetic carbon was shunted to the parasitic fungi, which can be grazed by zooplankton, thereby accelerating carbon transfer to higher trophic levels and bypassing the microbial loop. The herein termed “fungal shunt” can thus significantly modify the fate of photosynthetic carbon and the nature of phytoplankton–bacteria interactions, with implications for diverse pelagic food webs and global biogeochemical cycles.

Parasitism is one of the most common consumer strategies on Earth (1–3). Recently, it has also been identified as one of the dominating interactions within the planktonic interactome (4, 5), and yet parasites remain poorly considered in analyses of trophic interactions and element cycling in aquatic systems (6, 7). The foundation of trophic interactions in plankton communities is set by single-cell phytoplankton, which contributes almost half of the world’s primary production (8). According to our common understanding, the newly fixed carbon (C) is channeled either through the microbial loop, classical food web, or viral shunt, which supports the growth of heterotrophic bacteria and nanoflagellates, zooplankton and higher trophic levels, or viruses, respectively (9). However, fungi, particularly fungal microparasites, are rarely considered as contributors to C and nutrient cycling, although they are present and active in diverse aquatic environments (10–12).Members of the fungal division Chytridiomycota, referred to as chytrids, can thrive as microparasites on phytoplankton cells in freshwater (11, 13, 14) and marine systems (15–17), infecting up to 90% of the phytoplankton host population (18–21). A recent concept, called mycoloop, describes parasitic chytrids as an integral part of aquatic food webs (22). Energy and organic matter are thereby transferred from large, often inedible phytoplankton to chytrid zoospores, which are consumed by zooplankton (23–27). Hence, parasitic chytrids establish a novel trophic link between phytoplankton and zooplankton. Our understanding of element cycling and microbial interactions during chytrid epidemics, however, remains sparse. For instance, the cell-to-cell C transfer from single phytoplankton cells to their directly associated chytrids has not been quantified to date. Moreover, the relationship between parasitic chytrids and heterotrophic bacteria is largely undescribed.Phytoplankton cells release substantial amounts of dissolved organic C (DOC) (28), whereby up to 50% of photosynthetic C is consumed as DOC by bacteria (29–32). Thus, bacterial communities are intimately linked to phytoplankton abundances and production (33). Phytoplankton–bacteria interactions are particularly strong within the phycosphere, the region immediately surrounding individual phytoplankton cells (33–35), where nutrient concentrations are several-folds higher compared to the ambient water (36, 37), and nutrient assimilation rates of phytoplankton-associated bacteria are at least twice as fast as those of their free-living counterparts (38, 39). Importantly, parasitic chytrids may distort these phytoplankton–bacteria interactions within and outside the phycosphere since they modulate substrate and nutrient availabilities and presumably also bacterial activity and community composition. The effects of this distortion are virtually unresolved, but the few available data indicate that chytrid infections alter the composition and concentration of DOC (40), while abundances of free-living bacteria increase (25, 40) or remain unchanged (24).To disentangle phytoplankton–fungi–bacteria interactions at a microspatial single-cell scale—the scale at which phytoplankton, fungi, and bacteria intimately interact—we used one of the few existing model pathosystems, composed of the freshwater diatom Asterionella formosa, the chytrid Rhizophydiales sp., and coenriched populations of heterotrophic bacteria. Our methodology included dual stable-isotope incubations (¹³C-bicarbonate and ¹⁵N-nitrate), single-cell–resolution secondary ion mass spectrometry (SIMS) (IMS 1280 and NanoSIMS 50L), 16S rRNA gene/16S rRNA sequencing, microscopy (e.g., fluorescence in situ hybridization [FISH]), and nutrient analyses. We particularly focused on the initial C transfer from the phytoplankton host to parasitic chytrids, which we term the “fungal shunt,” as part of the mycoloop. The objectives were twofold: 1) quantifying the transfer of photosynthetic C from phytoplankton cells to infectious chytrids, cell-associated bacteria, and free-living bacteria and 2) characterizing the effect of parasitic fungi on bacterial populations, considering bacterial abundances, bacterial–diatom attachment, single-cell activity rates, and community composition. The obtained data challenge the common perception of aquatic microbial food webs by demonstrating the significant role that parasitic fungi can play in microbial community structure, interactions, and element cycling during phytoplankton growth.     

Prostate cancer marker panel with single cell sensitivity in urine

Background

Over one million men undergo prostate biopsies annually in the United States, a majority of whom due to elevated serum PSA. More than half of the biopsies turn out to be negative for prostate cancer (CaP). The limitations of both the PSA test and the biopsy procedure have led to the development for more precise CaP detection assays in urine (e.g., PCA3, TMPRSS2‐ERG) or blood (e.g., PHI, 4K). Here, we describe the development and evaluation of the Urine CaP Marker Panel (UCMP) assay for sensitive and reproducible detection of CaP cells in post‐digital rectal examination (post‐DRE) urine.

Methods

The cellular content of the post‐DRE urine was captured on a translucent filter membrane, which is placed on Cytoclear slides for direct evaluation by microscopy and immuno‐cytochemistry (ICC). Cells captured on the membrane were assayed for PSA and Prostein expression to identify prostate epithelial cells, and for ERG and AMACR to identify prostate tumor cells. Immunostained cells were analyzed for quantitative and qualitative features and correlated with biopsy positive and negative status for malignancy.

Results

The assay was optimized for single cell capture sensitivity and downstream evaluations by spiking a known number of cells from established CaP cell lines, LNCaP and VCaP, into pre‐cleared control urine. The cells captured from the post‐DRE urine of subjects, obtained prior to biopsy procedure, were co‐stained for ERG, AMACR (CaP specific), and Prostein or PSA (prostate epithelium specific) rendering a whole cell based analysis and characterization. A feasibility cohort of 63 post‐DRE urine specimens was assessed. Comparison of the UCMP results with blinded biopsy results showed an assay sensitivity of 64% (16 of 25) and a specificity of 68.8% (22 of 32) for CaP detection by biopsy.

Conclusions

This pilot study assessing a minimally invasive CaP detection assay with single cell sensitivity cell‐capture and characterization from the post‐DRE urine holds promise for further development of this novel assay platform. Prostate 75: 969–975, 2015. © 2015 The Authors. The Prostate, published by Wiley Periodicals, Inc.     

Need for permanent pacemaker implantation following implantation of the rapid deployment valve in combined procedures: a single centre cohort study

BackgroundRapid deployment aortic valves may interfere with the cardiac conduction system. We investigated the need for permanent pacemaker implantation (PPI) following the implantation of Edwards INTUITY valve (Edwards Lifesciences, Irvine, CA).MethodsOne hundred twenty patients underwent aortic valve replacement (AVR) with the INTUITY valve in a combined procedure at the German Heart Centre Munich between April 2016 and December 2019. Twenty-four patients with prior PPI or concomitant ablation procedures (24/120, 20%) were excluded. Patient-specific, procedural and post-procedural outcomes were assessed in the remaining 96 cases.ResultsAVR was successful in all cases. Seventy-four percent of the study population were men. Mean age was 69.5±7.6 years. EuroSCORE II was 3.2±2.9. Forty-six patients (46/96, 47.9%) presented with pre-operative conduction disorders, right bundle branch block (RBBB) (17/96, 17.7%) and first-degree or second degree atrio-ventricular block (AVB) (18/96, 18.8%), in particular. In total, 9 patients (9/96, 9.4%) underwent PPI. PPI was required in 3 patients (3/50, 6.0%) who did not have a pre-existing conduction disorder due to new high degree AVB. 6 patients with pre-operative conduction disorders (6/46, 13%) needed PPI. Timing of PPI was 5.2±1.5 days (median 5). Independent predictors of PPI were preoperative RBBB [odds ratio (OR) =4.554, P=0.049] and large valve size (#27) (OR =5.527, P=0.031).ConclusionsThe analysis of the data collected enabled us to identify patient factors associated with higher risk for post-operative PPI following AVR with the INTUITY valve. Patient factors associated with post-operative PPI, were RBBB and large valve size. These patients should be closely monitored following the procedure, in particular.