Urologische Klinik München

Abstract Functional genetic redundancy (FGR) within gene families limits the discovery of gene function in plants because single-gene perturbations often fail to produce informative phenotypes. Artificial microRNAs (amiRNAs) provide a strategy to silence multiple related genes simultaneously. However, the existing amiRNA-based libraries used for genetic gene function discovery in plants do not account for the subcellular localization of gene products, which can lead to pleiotropic or difficult-to-interpret phenotypes. Plastids are essential plant cell organelles that integrate central metabolic and signaling processes, including photosynthesis, hormone biosynthesis, and environmental responses. Here we introduce pamiR , a plastid-targeted amiRNA library designed to enable organelle-specific gene function discovery in Arabidopsis thaliana . Using plastid proteomic datasets, we identified high-confidence plastid-localized proteins and designed amiRNAs to target their gene(s) (families) minimizing FGR. This amiRNA library was introduced in a vector with fluorescence-accumulating seed technology enabling rapid, herbicide-free selection and screening in the first generation. Validation by next-generation sequencing, confirmed high representation and uniform distribution of amiRNAs within pamiR . Proof-of-concept screens recovered mutants affecting known and additional candidate genes involved in photosynthesis and abscisic acid biosynthesis. Therefore, the pamiR library provides a fast platform for plastid-focused genetic screens that is compatible with existing mutant collections. One-sentence summary The plastid amiRNA ( pamiR ) library enables organelle-specific forward genetics without functional genetic redundancy.

Genomic sequencing leaves >50% of dystonia-affected individuals without a diagnosis. Where DNA-oriented approaches remain insufficient, integrating multiomics is essential to advance genome interpretation. Herein, we incorporated RNA sequencing (RNA-seq) data from 167 patients with dystonia across a range of ages and presentations. We leveraged an RNA-seq analysis pipeline, focused on the identification of expression and splicing aberrations, on RNA-seq from skin biopsies. The recruited patients had early-onset dystonia in 85.0%, non-focal dystonia in 92.2%, and coexisting features in 76.0%. Thirty-six patient samples with pre-identified variants (36/167, 21.6%) and 131 samples with no previously prioritized diagnostic candidates from genomic sequencing (131/167, 78.4%) were evaluated. We found that >80% of dystonia-associated genes were detected by fibroblast RNA-seq. Expression and splicing aberration analyses produced a manageable number of significant RNA defects affecting dystonia-associated genes. The approach was especially successful in validating pathogenic effects of loss-of-function variants, with disease-relevant RNA-underexpression detected for 66.7% (10/15). Studying aberrant expression and splicing in the context of other pre-identified variant types yielded relevant results in 28.6% (6/21 samples). We obtained a 6.9% (9/131) diagnostic uplift for patients without prior candidates, all of whom exhibited combined dystonia with autosomal recessive inheritance. The new diagnoses from RNA-seq and genomic reanalysis were based on previously neglected splice-region (3/9) and deep(er) intronic (6/9) variants. For the observed events, integration of new machine-learning scores predicted corresponding aberrant gene expression in the brain. Fibroblast-based RNA-seq in our selected cohort improved variant interpretation and offered a modest yield in patients without prior candidate variants. ANN NEUROL 2026.

We hypothesized that B-GATA family transcription factors have important roles in growth regulation in moss. We analyzed B-GATA family transcription factor mutants from Physcomitrium patens and Arabidopsis thaliana to assess growth, gene expression, and cytokinin-related processes under varying nitrogen conditions. We found that nitrogen-dependent growth and transcriptional regulation are strongly impaired in mutants from Physcomitrium and Arabidopsis. We detected altered cytokinin homeostasis or signaling in the mutants, linking hormonal imbalance to growth and transcription defects. We demonstrated a conserved, critical role of B-GATAs in plant nitrogen-responsive growth. Results suggest that B-GATAs influence nitrogen-regulated transcription downstream from cytokinin, supporting an ancient, evolutionarily conserved mechanism connecting nutrient signaling to growth. We provided experimental evidence for the long-speculated but as-yet not demonstrated role of GATA transcription factors in nitrogen-dependent growth in land plants.

A secondary analysis of the PROPEL study (gov-Identifier: NCT00638235) data was conducted to compare symptom prevalence preoperatively vs. 6months after transvaginal prolapse repair with Elevate anterior and/or posterior. Symptoms were assessed with the pelvic floor distress inventory questionnaire (PFDI). Two hundred seventy-seven women with symptomatic II-IV stage POP underwent mesh-supported vaginal sacrospinous ligament fixation. Of these women, 187 (67%) reported at least one pain symptom of moderate or quite a bit severity preoperatively (anterior n = 105, visceral n = 129, posterior n = 122). Of these women reporting pain, approximately 40-64% had coexisting symptoms of urinary urgency, daytime urinary urgency, urinary urgency-incontinence, and nocturia of moderate or quite a bit severity. A smaller part reported coexisting symptoms of underactive bladder (UAB), fecal incontinence (FI), and/or obstructive defecation (OD). Sixmonths postoperatively, a significant reduction in the prevalence of almost all symptoms was observed. Cure rates for symptoms of overactive and underactive bladder were 65-85%, for symptoms of FI and OD 51-71%, and 58% (posterior), 85% (visceral), and 82% (anterior) for pain, respectively. The coexistence of bothersome pain, bladder, and bowel symptoms is common in women with POP. Therefore, POP should always be ruled out as a differential diagnosis before classifying the symptoms as interstitial cystitis/bladder pain syndrome. Women with POP and the co-existing symptoms described should be advised that surgical POP repair can resolve these symptoms in a high percentage.

In asymptomatic Seropositive/ACPA positive Rheumatoid Arthritis (RA) patients, we used a novel screening protocol for early subclinical detection of interstitial lung disease (ILD) based on lung function testing (PFT) and lung ultrasound (LUS). (Reichenberger et al BMJ Respir Med 2024). In a subset of patients, thoracic CT scan (TCT) has been performed in addition. We investigated 202 patients (76% female, mean age 60 +/- 12 years, 43% active or previous smokers) without symptoms of pulmonary disease. By means of LUS we found ILD suspicious changes in 56 patients (28%). A reduced DLCOc-SB</=80% were detected in 95 patients (47%), and a FVC </= 80% predicted were present in 30 patients (15%). In 31 patients (15%) early ILD was suspected based on reduced DLCOc-SB </=80% pred and noticeable changes in LUS. Patients with suspected ILD changes were significantly older, were more often active or ex smokers, had a significantly lower FVC and diffusion capacity, a lower VO2 at peak exercise and AT, a higher EQCO2 slope but no difference in disease duration of RA, disease activity of RA (DAS28-CRP Score), CCP titer, and biometric parameters. A TCT was performed in 42 patients, among them 14 with early signs of ILD. <table-wrap><object-id>erj;66/suppl_69/OA6453/TB1</object-id><object-id>T1</object-id><object-id>TB1</object-id><table><colgroup><col></col><col></col><col></col><col></col><col></col><col></col></colgroup><tbody><tr><td>Suspected ILD by Means of</td><td>ILD on TCT (n=14)</td><td>no ILD on TCT (n=28)</td><td>sens</td><td>spez</td><td>npv</td></tr><tr><td>DLCOc<80%</td><td>10</td><td>25</td><td>0,71</td><td>0,11</td><td>0,11</td></tr><tr><td>LUS positive</td><td>13</td><td>11</td><td>0,93</td><td>0,61</td><td>0,98</td></tr><tr><td>combination DLCO and LUS</td><td>12</td><td>17</td><td>0,86</td><td>0,61</td><td>0,96</td></tr></tbody></table></table-wrap> <bold>Conclusion:</bold> In asymptomatic RA-patients, signs of ILD based on PFT and LUS were consistently found in 15 % of patients. In a subset of 42 Patients who underwent TCT, LUS/PFT criteria has a NPV of 96% to exclude subclinical ILD

ABSTRACT Heterobranchia is a subclass of Gastropoda for which phylogenetic analyses based on mitochondrial gene sequences have not been entirely successful. Here, we report a complete mitochondrial genome of an ellobiid snail, Marinula pepita (Euthyneura: Eupulmonata: Ellobioidea), and its wholesale gene rearrangements from the presumed ancestral order for Heterobranchia. Our phylogenetic reconstruction based on this new genome and the available mitogenomic sequences recovered two sister clades in the superfamily Ellobioidea. Six genera with the ancestral gene order and short branches formed one of the clades (Trimusculus, Carychium, Ovatella, Ellobium, Leucophytia and Auriculastra). Four ellobiid genera with rearranged gene orders and long branches formed the other clade (Marinula, Pedipes, Melampus and Myosotella), regardless of their subfamilial assignments, thereby strongly questioning this topology [long branch attraction (LBA)]. A separate phylogenetic tree inferred from near-complete sequences of the nuclear 18S and 28S rRNA genes, on the other hand, was largely consistent with the conventional, morphology-based classification although suggesting the need of a new subfamily for Marinula. Mitochondrial sequences can be useful for inferring shallow divergences among certain ellobioid genera and species, provided that particular attention is paid to avoid LBA artefacts.

BackgroundBlock-face scanning electron microscopy has opened a new era of connectomics research, in which it is possible to make dense reconstructions of all cells in a clipping of a neuronal network, such as the retina, resolving synaptic contacts. Anchovies, exceptionally abundant marine teleosts, have retinae with regions for triple cone-based color vision and a region with specialized cone photoreceptors, so-called polycones, made of long and short cones with axially oriented outer segment lamellae for polarization contrast vision. This modality, discovered in the 1970s, is unique in vertebrates, but the neural wiring for contrast generation in deeper retinal layers is unknown so far.ResultsTo elucidate the retinal connectomics of the European anchovy Engraulis encrasicolus (Linnaeus, 1758), in a first project, we investigated the shapes and cone-specific wiring rules of 3 horizontal cell types using volume electron microscopy and subsequent computer-aided reconstruction: H1 cells contact both cone types of the polycone, H2 cells contact only the short cones, and H3 cells are exclusively connected to rods. In addition, a distinctive double band of Müller fibers and a layer of H1 axon terminals were structurally clarified.ConclusionsThe findings suggest that (1) the monochromatic polarization contrast system based on fine structure specializations in the outer retina is connected to an inherited (bichromatic) color contrast mechanism in the inner retina, (2) the anchovy polycones arose from red (now long) and green (now short) cones, and (3) the blue single cones disappeared in the relevant retinal region.

Potatoes were first brought to Europe in the sixteenth century1,2. Two hundred years later, one of the species had become one of the most important food sources across the entire continent and, later, even the entire world3. However, its highly heterozygous, autotetraploid genome has complicated its improvement since then4, 5, 6–7. Here we present the pan-genome of European potatoes generated from phased genome assemblies of ten historical potato cultivars, which includes approximately 85% of all haplotypes segregating in Europe. Sequence diversity between the haplotypes was extremely high (for example, 20× higher than in humans), owing to numerous introgressions from wild potato species. By contrast, haplotype diversity was very low, in agreement with the population bottlenecks caused by domestication and transition to Europe. To illustrate a practical application of the pan-genome, we converted it into a haplotype graph and used it to generate phased, megabase-scale pseudo-genome assemblies of commercial potatoes (including the famous French fries potato ‘Russet Burbank’) using cost-efficient short reads only. In summary, we present a nearly complete pan-genome of autotetraploid European potato, we describe extraordinarily high sequence diversity in a domesticated crop, and we outline how this resource might be used to accelerate genomics-assisted breeding and research.