Background:Understanding the phenotypic spectrum of disease-associated genes is essential for accurate diagnosis and targeted therapy. FRMPD4 (FERM and PDZ Domain Containing 4) has previously been associated with intellectual disability and epilepsy. However, its potential role in non-syndromic hearing loss has not been explored. Methods:We performed genetic analysis in two unrelated families presenting with non-syndromic sensorineural hearing loss, identifying maternally inherited missense variants in FRMPD4. Clinical phenotyping included audiological assessment and evaluation for neurodevelopmental involvement. Cross-species expression analyses were conducted in Drosophila, zebrafish, and mouse. Functional characterization included quantitative evaluation of sound-evoked responses in Drosophila nicht gut hörend (ngh) mutants, assessment of neuronal development and acoustic startle responses in zebrafish loss of function models, and morphological cochlear analyses with auditory brainstem response measurements in knockout mice. Results:Three affected males from two unrelated families presented with prelingual, bilaterally symmetrical sensorineural hearing loss, with confirmed congenital onset in one individual and no evidence of neurodevelopmental abnormalities. Cross-species analyses demonstrated evolutionarily conserved expression of FRMPD4 in auditory structures. In Drosophila, quantitative analysis of sound-evoked responses in ngh mutants revealed impaired auditory function. Zebrafish loss of function models exhibited reduced neuronal populations in the otic vesicle and posterior lateral line, abnormal neuromast development, and diminished acoustic startle responses. In mice, Frmpd4 knockout resulted in high-frequency hearing loss and cochlear abnormalities consistent with the human phenotype. Conclusions:Our findings expand the phenotypic spectrum of FRMPD4 to include non-syndromic sensorineural hearing loss and establish its evolutionarily conserved role in auditory function. These results have direct implications for genetic diagnosis and variant interpretation in patients with hearing loss.

Little is known about the importance of different tree species in maintaining diversity and function. To address this, we fogged 99 trees (30 Fraxinus excelsior, 30 F. pennsylvanica, 21 Quercus robur, 12 Ulmus laevis and 6 Tilia cordata) in Elbe floodplain forests, resulting in 182,179 arthropods including 25,861 (14.2%) beetles. Ninety-two percent of all beetles and 65% of all species were canopy associated. Tourist beetles of the ground contributed 35% to the arboreal richness but only 8% to overall abundance, suggesting little influence on trophic interactions. The highest alpha and beta diversity was recorded on Fraxinus trees, mainly due to the high number of tourists in low abundance. On the contrary, Quercus or Ulmus were characterised by large numbers of associated species. The beetle communities, and even more so the entire communities of all orders, were dominated by predators, suggesting strong top-down control of herbivore populations. Phytophages were significantly more abundant on Quercus and Ulmus, while they represented less than 10% on the ashes. F. excelsior is a keystone species, and this includes the ability to maintain biodiversity and communities with different functional profiles. As a result of ash dieback and drought, F. excelsior has been severely reduced in these forests. Our extensive data show that F. excelsior and F. pennsylvanica are similar in many ecological features due to their taxonomic relationship, resulting in a similar diversity and functional composition of arthropod communities, indicating that F. pennsylvanica can at least partially fill the ecological role of the common ash.

The sperm epigenome is thought to affect the developmental programming of the resulting embryo, influencing health and disease in later life. Age-related methylation changes in the sperm of old fathers may mediate the increased risks for reproductive and offspring medical problems. The impact of paternal age on sperm methylation has been extensively studied in humans and, to a lesser extent, in rodents and cattle. Here, we performed a comparative analysis of paternal age effects on protein-coding genes in the human and marmoset sperm methylomes. The marmoset has gained growing importance as a non-human primate model of aging and age-related diseases. Using reduced representation bisulfite sequencing, we identified age-related differentially methylated transcription start site (ageTSS) regions in 204 marmoset and 27 human genes. The direction of methylation changes was the opposite, increasing with age in marmosets and decreasing in humans. None of the identified ageTSS was differentially methylated in both species. Although the average methylation levels of all TSS regions were highly correlated between marmosets and humans, with the majority of TSS being hypomethylated in sperm, more than 300 protein-coding genes were endowed with species-specifically (hypo)methylated TSS. Several genes of the glycosphingolipid (GSL) biosynthesis pathway, which plays a role in embryonic stem cell differentiation and regulation of development, were hypomethylated (<5%) in human and fully methylated (>95%) in marmoset sperm. The expression levels and patterns of defined sets of GSL genes differed considerably between human and marmoset pre-implantation embryo stages and blastocyst tissues, respectively.

For decades, frustrated quantum magnets have been a seed for scientific progress and innovation in condensed matter. As much as the numerical tools for low-dimensional quantum magnetism have thrived and improved in recent years due to breakthroughs inspired by quantum information and quantum computation, higher-dimensional quantum magnetism can be considered as the final frontier, where strong quantum entanglement, multiple ordering channels, and manifold ways of paramagnetism culminate. At the same time, efforts in crystal synthesis have induced a significant increase in the number of tangible frustrated magnets which are generically three-dimensional in nature, creating an urgent need for quantitative theoretical modeling. We review the pseudo-fermion (PF) and pseudo-Majorana (PM) functional renormalization group (FRG) and their specific ability to address higher-dimensional frustrated quantum magnetism. First developed more than a decade ago, the PFFRG interprets a Heisenberg model Hamiltonian in terms of Abrikosov pseudofermions, which is then treated in a diagrammatic resummation scheme formulated as a renormalization group flow of $m$-particle pseudofermion vertices. The article reviews the state of the art of PFFRG and PMFRG and discusses their application to exemplary domains of frustrated magnetism, but most importantly, it makes the algorithmic and implementation details of these methods accessible to everyone. By thus lowering the entry barrier to their application, we hope that this review will contribute towards establishing PFFRG and PMFRG as the numerical methods for addressing frustrated quantum magnetism in higher spatial dimensions.

As a highly frustrated model Hamiltonian with an exact dimer ground state, the Heisenberg antiferromagnet on the maple leaf lattice is of high theoretical interest, and a material realization is intensely sought after. We determine the magnetic Hamiltonian of the copper mineral bluebellite using density functional theory based energy mapping. As a consequence of the significant distortion of the spin $S=1/2$ maple leaf lattice, we find two of the five distinct nearest neighbor couplings to be ferromagnetic. Solution of this Hamiltonian with density matrix renormalization group calculations points us to the surprising insight that this particular imperfect maple leaf lattice, due to the strongly ferromagnetic Cu$^{2+}$ dimer, realizes an effective $S=1$ breathing kagome Hamiltonian. In fact, this is another highly interesting Hamiltonian which has rarely been realized in materials. Analysis of the effective model within a bond-operator formalism allows us to identify a valence bond solid ground state and to extract thermodynamic quantities using a low-energy bosonic mean-field theory. We resolve the puzzle of the apparent one-dimensional character of bluebellite as our calculated specific heat has a Bonner-Fisher-like shape, in good agreement with experiment.

Optimal stomatal regulation is important for plant adaptation to changing environmental conditions and for maintaining crop yield. The guard cell signal γ-aminobutyric acid (GABA) is produced from glutamate by glutamate decarboxylase (GAD) during a reaction that generates CO2 as a by-product. Here, we investigated a putative connection between GABA signalling and the more clearly defined CO2 signalling pathway in guard cells. The GABA-deficient mutant Arabidopsis lines gad2-1, gad2-2, and gad1/2/4/5 were examined for stomatal sensitivity to various CO2 concentrations. Our findings show a phenotypical discrepancy between the allelic mutant lines gad2-1 and gad2-2-a weakened CO2 response in gad2-1 (GABI_474_E05) in contrast to a wild-type response in gad2-2 (SALK_028819) and gad1/2/4/5. Through transcriptomic and genomic investigation, we traced the response of gad2-1 to a deletion of full-length Mitogen-activated protein kinase 12 (MPK12) in the GABI-KAT line, thereafter renamed as gad2-1*. Guard cell-specific complementation of MPK12 in gad2-1* restored the wild-type CO2 phenotype, which confirms the proposed importance of MPK12 in CO2 sensitivity. Additionally, we found that stomatal opening under low atmospheric CO2 occurs independently of the GABA-modulated opening channel ALUMINIUM-ACTIVATED MALATE TRANSPORTER 9 (ALMT9). Our results demonstrate that GABA has a role in modulating the rate of stomatal opening and closing, but not in response to CO2per se.

Copy number variations (CNVs) of the KITLG gene seem to be involved in the oncogenesis of digital squamous cell carcinoma (dSCC). The aims of this study were (1) to investigate KITLG CNV in giant (GS), standard (SS), and miniature (MS) schnauzers and (2) to compare KITLG CNV between black GS with and without dSCC. Blood samples from black GS (22 with and 17 without dSCC), black SS (18 with and 4 without dSSC; 5 unknown), and 50 MS (unknown dSSC status and coat colour) were analysed by digital droplet PCR. The results are that (1) most dogs had a copy number (CN) value > 4 (range 2.5–7.6) with no significant differences between GS, SS, and MS, and (2) the CN value in black GS with dSCC was significantly higher than in those without dSCC (p = 0.02). CN values > 5.8 indicate a significantly increased risk for dSCC, while CN values < 4.7 suggest a reduced risk for dSCC (grey area: 4.7–5.8). Diagnostic testing for KITLG CNV may sensitise owners to the individual risk of their black GS for dSCC. Further studies should investigate the relevance of KITLG CNV in SS and the protective effects in MS, who rarely suffer from dSCC.

We observe a localized cnoidal (LCn) state in an electric circuit network. Its formation derives from the interplay of nonlinearity and the topology inherent to a Su-Schrieffer-Heeger (SSH) chain of inductors. Varicap diodes act as voltage-dependent capacitors, and create a nonlinear on-site potential. For a sinusoidal voltage excitation around midgap frequency, we show that the voltage response in the nonlinear SSH circuit follows the Korteweg-de Vries equation. The topological SSH boundary state, which relates to a midgap impedance peak in the linearized limit is distorted into the LCn state in the nonlinear regime, where the cnoidal eccentricity decreases from edge to bulk.

Advanced paternal age is associated with increased risks for reproductive and offspring medical problems.Accumulating evidence suggests age-related changes in the sperm epigenome as one underlying mechanism.Using reduced representation bisulfite sequencing on 73 sperm samples of males attending a fertility center, we identified 1,162 (74%) regions which were significantly (FDR-adjusted) hypomethylated and 403 regions (26%) being hypermethylated with age.There were no significant correlations with paternal BMI, semen quality, or ART outcome.The majority (1,152 of 1,565; 74%) of age-related differentially methylated regions (ageDMRs) were located within genic regions, including 1,002 genes with symbols.Hypomethylated ageDMRs were closer to transcription start sites than hypermethylated DMRs, half of which reside in gene-distal regions.In this and conceptually related genome-wide studies, so far 2,355 genes have been reported with significant sperm ageDMRs, however most (90%) of them in only one study.The 241 genes which have been replicated at least once showed significant functional enrichments in 41 biological processes associated with development and the nervous system and in 10 cellular components associated with synapses and neurons.This supports the hypothesis that paternal age effects on the sperm methylome affect offspring behaviour and neurodevelopment.It is interesting to note that sperm ageDMRs were not randomly distributed throughout the human genome; chromosome 19 showed a highly significant twofold enrichment with sperm ageDMRs.Although the high gene density and CpG content have been conserved, the orthologous marmoset chromosome 22 did not appear to exhibit an increased regulatory potential by age-related DNA methylation changes.

The square-kagome lattice Heisenberg antiferromagnet is a highly frustrated Hamiltonian whose material realizations have been scarce. We theoretically investigate the recently synthesized Na6Cu7BiO4(PO)4)4Cl3 where a Cu2+ spin-1/2 square-kagome lattice (with a six site unit cell) is decorated by a seventh magnetic site alternatingly above and below the layers. The material does not show any sign of long-range magnetic order down to 50 mK despite a Curie-Weiss temperature of -212 K indicating a quantum paramagnetic phase. Our DFT energy mapping elicits a purely antiferromagnetic Hamiltonian that features longer range exchange interactions beyond the pure square-kagome model and, importantly, we find the seventh site to be strongly coupled to the plane. We combine two variational Monte Carlo approaches, pseudofermion/Majorana functional renormalization group and Schwinger-Boson mean field calculations to show that the complex Hamiltonian of Na6Cu7BiO4(PO)4)4Cl3 still features a nonmagnetic ground state. We explain how the seventh Cu2+ site actually aids the stabilization of the disordered state. We predict static and dynamic spin structure factors to guide future neutron scattering experiments.

We analyze the quantum antiferromagnet on the maple leaf lattice in the presence of a magnetic field. Starting from its exact dimer ground state and for a magnetic field strength of the order of the local dimer spin exchange coupling, we perform a strong coupling expansion and extract an effective hardcore boson model. The interplay of effective many-body interactions, suppressed single-particle dynamics, and correlated hopping gives way to an intriguing series of superfluid to insulator transitions which correspond to magnetization plateaux in terms of the maple leaf spin degrees of freedom. While we find plateaux at intermediate magnetization to be dominated by bosonic density wave order, we conjecture plateau formation from multi-boson bound states due to correlated hopping for lower magnetization.

In schnauzers, a breed predisposition to squamous cell carcinoma of the digit (dSCC) is well known. The aim of this study was to compare the clinical and macroscopic findings of dSCCs in giant (GSs), standard (SSs), and miniature schnauzers (MSs). Methods: Pathology reports of 478 dSCCs from 417 schnauzers (227 GSs, 174 SSs, and 16 MSs) were retrospectively evaluated. Results: The MSs were older than the SSs and GSs (p ≤ 0.01). The male GSs were predisposed to dSCC (p < 0.05). In the GSs, the nodular dSCCs were larger than in the MSs (p ≤ 0.05) and SSs (p ≤ 0.001). The digital SCCs were mostly diagnosed at the forelimbs, especially at digits 1, 2, and 5. At the hindlimbs, the affected toes differed between the GSs and SSs. Multiple dSCCs were more common in SSs than in GSs (p = 0.003). If dSCC was the cause of death, the survival time was shorter than in dogs dying from other diseases (p = 0.004). Metastases occurred in 20% of the cases and led to a significantly shorter survival time in both the GSs and SSs (p < 0.001). Conclusions: The results showed various differences in the dSCC depending on the size variant of the schnauzer.

Dark-haired dogs are predisposed to the development of digital squamous cell carcinoma (DSCC). This may potentially suggest an underlying genetic predisposition not yet completely elucidated. Some authors have suggested a potential correlation between the number of copies KIT Ligand (KITLG) and the predisposition of dogs to DSCC, containing a higher number of copies in those affected by the neoplasm. In this study, the aim was to evaluate a potential correlation between the number of copies of the KITLG and the histological grade of malignancy in dogs with DSCC. For this, 72 paraffin-embedded DSCCs with paired whole blood samples of 70 different dogs were included and grouped according to their haircoat color as follow: Group 0/unknown haircoat color (n = 11); Group 1.a/black non-Schnauzers (n = 15); group 1.b/black Schnauzers (n = 33); group 1.c/black and tan dogs (n = 7); group 2/tan animals (n = 4). The DSCCs were histologically graded. Additionally, KITLG Copy Number Variation (CNV) was determined by ddPCR. A significant correlation was observed between KITLG copy number and the histological grade and score value. This finding may suggest a possible factor for the development of canine DSCC, thus potentially having an impact on personalized veterinary oncological strategies and breeding programs.

OBJECTIVES:Preoperative identification of malignant adrenal tumors is challenging. 24-h urinary steroid profiling by LC-MS/MS and machine learning has demonstrated high diagnostic power, but the unavailability of bioinformatic models for public use has limited its routine application. We here aimed to increase usability with a novel classification model for the differentiation of adrenocortical adenoma (ACA) and adrenocortical carcinoma (ACC). METHODS:Eleven steroids (5-pregnenetriol, dehydroepiandrosterone, cortisone, cortisol, α-cortolone, tetrahydro-11-deoxycortisol, etiocholanolone, pregnenolone, pregnanetriol, pregnanediol, and 5-pregnenediol) were quantified by LC-MS/MS in 24-h urine samples from 352 patients with adrenal tumor (281 ACA, 71 ACC). Random forest modelling and decision tree algorithms were applied in training (n = 188) and test sets (n = 80) and independently validated in 84 patients with paired 24-h and spot urine. RESULTS:After examining different models, a decision tree using excretions of only 5-pregnenetriol and tetrahydro-11-deoxycortisol classified three groups with low, intermediate, and high risk for malignancy. 148/217 ACA were classified as being at low, 67 intermediate, and 2 high risk of malignancy. Conversely, none of the ACC demonstrated a low-risk profile leading to a negative predictive value of 100% for malignancy. In the independent validation cohort, the negative predictive value was again 100% in both 24-h urine and spot urine with a positive predictive value of 87.5% and 86.7%, respectively. CONCLUSIONS:This simplified LC-MS/MS-based classification model using 24-h-urine provided excellent results for exclusion of ACC and can help to avoid unnecessary surgeries. Analysis of spot urine led to similarly satisfactory results suggesting that cumbersome 24-h urine collection might be dispensable after future validation.

A prerequisite for sustainable forest management is knowing the total diversity and how management affects forests. Both are poorly studied and relate to canopy diversity and comparison with primary forests. From 2001–2004, we fogged beetles from oaks in primary and disturbed, managed sites in Białowieża (Eastern Poland) and also in distant age-class forests. Using a machine learning (ML) method (elastic net), we identified a beetle signature based on the species abundance distribution to distinguish these forest types. The beetle communities from 2001 served as training data, with 21 signature species correctly assigning the oaks to primary and different managed forests. However, the predictive quality of the signature decreased with each year due to high spatio-temporal heterogeneity and beta diversity. To improve the power of the signature, we combined the data from all years to calculate a more general model. Due to its greater complexity, this model identified 60 species that correctly classified both the studied forests and foreign forests in Central Europe, increasing the possibility of a general classification. Further research is needed to determine whether it is possible to establish a general signature-based index on a large number of samples from different years and forest types.

Grading, immunohistochemistry and c-kit mutation status are criteria for assessing the prognosis and therapeutic options of canine cutaneous mast cell tumours (MCTs). As a subset, canine digital MCTs have rarely been explored in this context. Therefore, in this retrospective study, 68 paraffin-embedded canine digital MCTs were analysed, and histological grading was assessed according to Patnaik and Kiupel. The immunohistochemical markers KIT and Ki67 were used, as well as polymerase chain reaction (PCR) for mutational screening in c-kit exons 8, 9, 11 and 14. Patnaik grading resulted in 22.1% grade I, 67.6% grade II and 10.3% grade III tumours. Some 86.8% of the digital MCTs were Kiupel low-grade. Aberrant KIT staining patterns II and III were found in 58.8%, and a count of more than 23 Ki67-positive cells in 52.3% of the cases. Both parameters were significantly associated with an internal tandem duplication (ITD) in c-kit exon 11 (12.7%). French Bulldogs, which tend to form well-differentiated cutaneous MCTs, had a higher proportion of digital high-grade MCTs and ITD in c-kit exon 11 compared with mongrels. Due to its retrospective nature, this study did not allow for an analysis of survival data. Nevertheless, it may contribute to the targeted characterisation of digital MCTs.

Although much is known about the ecology and functional importance of canopy arthropods in temperate forests, few studies have tried to assess the overall diversity and investigate the composition and dynamics of tree-specific communities. This has impeded a deeper understanding of the functioning of forests, and of how to maintain system services. Here, we present the first comprehensive data of whole arthropod communities, collected by insecticidal knockdown (fogging) from 1159 trees in 18 study areas in Central Europe during the last 25 years. The data includes 3,253,591 arthropods from 32 taxa (order, suborder, family) collected on 24 tree species from 18 genera. Fogging collects free-living, ectophytic arthropods in approximately the same number as they occur in the trees. To our knowledge, these are the most comprehensive data available today on the taxonomic composition of arboreal fauna. Assigning all arthropods to their feeding guild provided a proxy of their functional importance. The data showed that the canopy communities were regularly structured, with a clear dominance hierarchy comprised of eight 'major taxa' that represented 87% of all arthropods. Despite significant differences in the proportions of taxa on deciduous and coniferous trees, the composition of the guilds was very similar. The individual tree genera, on the other hand, showed significant differences in guild composition, especially when different study areas and years were compared, whereas tree-specific traits, such as tree height, girth in breast height or leaf cover, explained little of the overall variance. On the ordinal level, guild composition also differed significantly between managed and primary forests, with a simultaneous low within-group variability, indicating that management is a key factor determining the distribution of biodiversity and guild composition.

Renormalization group methods are well-established tools for the (numerical) investigation of the low-energy properties of correlated quantum many-body systems, allowing us to capture their scale-dependent nature. The functional renormalization group (FRG) allows us to continuously evolve a microscopic model action to an effective low-energy action as a function of decreasing energy scales via an exact functional flow equation, which is then approximated by some truncation scheme to facilitate computation. Here, we report on our implementation of multiloop FRG, an extended truncation scheme recently developed for electronic FRG calculations, within the pseudofermion functional renormalization group (pf-FRG) framework for interacting quantum spin systems. We discuss in detail the conceptual intricacies of the flow equations generated by the multiloop truncation, as well as essential refinements to the integration scheme for the resulting integrodifferential equations. To benchmark our approach, we analyze antiferromagnetic Heisenberg models on the pyrochlore, simple cubic, and face-centered cubic lattice, discussing the convergence of physical observables for higher-loop calculations and comparing with existing results where available. Combined, these methodological refinements systematically improve the pf-FRG approach to one of the numerical tools of choice when exploring frustrated quantum magnetism in higher spatial dimensions.

We studied the impact of the neophyte tree Fraxinus pennsylvanica on the diversity of beetles in floodplain forests along the river Elbe in Germany in 2016, 2017 and in 2020, where 80% of all Fraxinus excelsior trees had died following severe droughts. Beetles were collected by insecticidal knock-down from 121 trees (64 F. excelsior and 57 F. pennsylvanica) and identified to 547 species in 15,214 specimens. The trees sampled in 2016 and 2017 showed no signs of drought stress or ash dieback and serve as a reference for the comparison with the 2020 fauna. The data proved that F. excelsior harbours the most diverse beetle community, which differed also significantly in guild composition from F. pennsylvanica. Triggered by extremely dry and long summer seasons, the 2020 ash dieback had profound and forest-wide impacts. Several endangered, red-listed beetle species of Saxonia Anhalt had increased in numbers and became secondary pests on F. excelsior. Diversity decreased whilst numbers of xylobionts increased on all trees, reaching 78% on F. excelsior. Proportions of xylobionts remained constant on F. pennsylvanica. Phytophages were almost absent from all trees, but mycetophages increased on F. pennsylvanica. Our data suggest that as a result of the dieback of F. excelsior the neophyte F. pennsylvanica might become a rescue species for the European Ash fauna, as it provides the second-best habitat. We show how difficult it is to assess the dynamics and the ecological impact of neophytes, especially under conditions similar to those projected by climate change models. The diversity and abundance of canopy arthropods demonstrates their importance in understanding forest functions and maintenance of ecosystem services, illustrating that their consideration is essential for forest adaptation to climate change.