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Abstract Zirconia-based shape memory ceramics (SMCs) exhibit anisotropic mechanical response when undergoing elastic deformations as well as during austenite–martensite phase transformation. This behavior results in different types of strain incompatibility at grain boundaries, which we study here using a micromechanical model. A single-crystal model is implemented to provide a full mechanistic three-dimensional description of the anisotropic elastic as well as martensitic transformation stress–strain response, including non-Schmid behavior caused by the significant volume change during martensitic transformation. This model was calibrated to and validated against compression tests of single-crystal zirconia micro-pillars conducted previously, and then used to model bi-crystals. Upon the introduction of a grain boundary, the simulation provides detailed information on the nucleation and evolution of martensite variants and stress distribution at grain boundaries. We identify bi-crystal configurations which result in very large stress concentrations at very low deformations due to elastic incompatibility, as well as others where the elastic incompatibility is relatively low and stress concentrations only occur at large transformation strains. We also show how this approach can be used to explore the misorientation space for quantifying the level of elastic and transformation incompatibility at SMCs grain boundaries. Graphical abstract Micromechanics models provide insights on grain boundary elastic and phase transformation strain incompatibility in shape memory zirconia

Studying states and state transitions in the brain is challenging due to the nonlinear, complex dynamics present. In this research, we analyse the brain’s response to non-invasive perturbations. Perturbation techniques offer a powerful method for studying complex dynamics [1], though their translation to human brain data is under-explored. This method involves applying small inputs, in this case via photic stimulation, to a system and measuring its response. Sensitivity to perturbations can forewarn a state transition [1], therefore biomarkers of the brain’s perturbation response or ‘cortical excitability’ could be used to indicate seizure transitions [2; 3; 4]. However, perturbing the brain often involves invasive intracranial surgeries [5] or expensive equipment as in transcranial magnetic stimulation (TMS) which is only accessible to a minority of patient groups [6; 7], or animal model studies [8]. Photic stimulation is a widely used diagnostic technique in epilepsy [9] that can be used as a non-invasive perturbation paradigm to probe brain dynamics during routine electroencephalography (EEG) studies in humans. This involves changing the frequency of strobing light, sometimes triggering a photo-paroxysmal response (PPR), which is an electrographic event that can be studied as a state transition to a seizure state. We investigate alterations in the response to these perturbations in patients with genetic generalised epilepsy (GGE), with (n=10) and without (n=10) PPR, and patients with psychogenic non-epileptic seizures (PNES; n=10), compared to resting controls (n=10). Metrics of EEG time-series data were evaluated as biomarkers of the perturbation response including variance, autocorrelation, and phase-based synchrony measures. We observed considerable differences in all group biomarker distributions during stimulation compared to controls. In particular, variance and autocorrelation demonstrated greater changes in epochs close to PPR transitions compared to earlier stimulation epochs. Comparison of PPR and spontaneous seizure morphology found them indistinguishable, suggesting PPR is a valid proxy for seizure dynamics. Also, as expected, posterior channels demonstrated the greatest change in synchrony measures, possibly reflecting underlying PPR pathophysiology mechanisms [9]. We clearly demonstrate observable changes at a group level in cortical excitability in epilepsy patients as a response to perturbation in EEG data. Our work re-frames photic stimulation as a non-invasive perturbation paradigm capable of inducing measurable changes to brain dynamics.

Clinical trials and proof-of-concept pre-clinical work for different cancers has shown relatively positive outcomes and tumour killing when p53, a well-established tumour suppressor, levels and function is restored. Human papillomavirus (HPV) driven cancers encode the E6 oncoprotein that degrades p53 to allow HPV-driven carcinogenic process to proceed. Indeed, there have been several attempts in the past to revive p53 function in HPV driven cancers by pharmacological and genetic means to increase p53 bioavailability. Here, we employed a CRISPR activation (CRISPRa) approach to overcome HPV-mediated silencing of p53 by hyper expressing the p53 promoter. Hyperexpression of p53 led to HPV positive (+) cervical cancer cell killing and reduced cell proliferation. Increasing p53 bioavailability may have promising potential as a therapy for HPV driven cancers.

Intergenerational Health Review (IHR) postulates that reminiscence of personal history is a reconciling process beneficial to individuals since it helps to find meaning in life and gain a better se...

Objective. Long-term electroencephalogram (EEG) recordings can aid diagnosis and management of various neurological conditions such as epilepsy. In this study we characterize the safety and stability of a clinical grade ring electrode arrays by analyzing EEG recordings, fluoroscopy, and computed tomography (CT) imaging with long-term implantation and histopathological tissue response. Approach. Seven animals were chronically implanted with EEG recording array consisting of four electrode contacts. Recordings were made bilaterally using a bipolar longitudinal montage. The array was connected to a fully implantable micro-processor controlled electronic device with two low-noise differential amplifiers and a transmitter-receiver coil. An external wearable was used to power, communicate with the implant via an inductive coil, and store the data. The sub-scalp electrode arrays were made using medical grade silicone and platinum. The electrode arrays were tunneled in the subgaleal cleavage plane between the periosteum and the overlying dermis. These were implanted for 3–7 months before euthanasia and histopathological assessment. EEG and impedance were recorded throughout the study. Main results. Impedance measurements remained low throughout the study for 11 of 12 channels over the recording period ranged from 3 to 5 months. There was also a steady amplitude of slow-wave EEG and chewing artifact (noise). The post-mortem CT and histopathology showed the electrodes remained in the subgaleal plane in 6 of 7 sheep. There was minimal inflammation with a thin fibrotic capsule that ranged from 4 to 101 μm. There was a variable fibrosis in the subgaleal plane extending from 210 to 3617 μm (S3–S7) due to surgical cleavage. One sheep had an inflammatory reaction due to electrode extrusion. The passive electrode array extraction force was around 1 N. Significance. Results show sub-scalp electrode placement was safe and stable for long term implantation. This is advantageous for diagnosis and management of neurological conditions where long-term, EEG monitoring is required.

Intergenerational study tourism (IST) is a conceptual model designed to promote cross-cultural, intergenerational (IG) encounters through travel and learning. IST is an area of IG practice that wou...

In the contexts of population aging and English as a second language (ESL) proficiency decline in Hong Kong, a 12 weeks program that matched 15 retired citizens with 30 secondary school children wh...

Malaria causes more than 200 million clinical illnesses and 45 million deaths every year, making mass production of an effective vaccine increasingly urgent. A Plasmodium falciparum sporozoites (PfSPZ) based vaccine has been proved to be a promising choice to defend against the malaria pandemic. However, large scale industrial production of PfSPZ vaccine is currently sub-optimal as the process for sporozoite extraction from salivary glands of infected mosquitoes is performed by manual microdissection, a relatively inefficient process requiring many hours of training. This paper reports continued progress in our development of a robotic system for automating the extraction of salivary glands. Compared to our previous versions, the structure of the new system is optimized to be more compact and allows different steps in workflow to be performed in parallel. We also report continued progress in developing key subsystem components. Finally, experiments show encouraging results with success rates of 90% in robotic mosquito manipulation, 95.6% in salivary gland extraction and 92.7% in debris disposal.

For idiopathic generalized epilepsies (IGE), brain network analysis is emerging as a biomarker for potential use in clinical care. To determine whether people with IGE show alterations in resting-state brain connectivity compared to healthy controls, and to quantify these differences, we conducted a systematic review and meta-analysis of EEG and magnetoencephalography (MEG) functional connectivity and network studies. The review was conducted according to PRISMA guidelines. Twenty-two studies were eligible for inclusion. Outcomes from individual studies supported hypotheses for interictal, resting-state brain connectivity alterations in IGE patients compared to healthy controls. In contrast, meta-analysis from six studies of common network metrics clustering coefficient, path length, mean degree and nodal strength showed no significant differences between IGE and control groups (effect sizes ranged from -0.151 -1.78). The null findings of the meta-analysis and the heterogeneity of the included studies highlights the importance of developing standardized, validated methodologies for future research. Network neuroscience has significant potential as both a diagnostic and prognostic biomarker in epilepsy, though individual variability in network dynamics needs to be better understood and accounted for.

To improve outdoor thermal environment and reduce indoor energy use, passive techniques including facade greenery have been suggested. In high-density cities like Hong Kong, buildings’ surface area is much greater than the roof and ground surface areas combined, offering a huge vertical surface platform for greening. However, scientific evidence to assert the thermal benefit from this greening option especially at neighborhood scale is still very few. Therefore, this study was designed to provide such evidence using results from validated ENVI-met model simulation. Thereafter, parametric study was conducted to investigate the quantity and location of facade greening required for potential air cooling and thermal comfort improvement of a neighborhood of varying densities. Model validation results revealed an acceptable modelling of facade surface temperature, air temperature, relative humidity and wall-emitted long-wave fluxes. From the parametric study, we found that 30–50% of facades in the high-density urban setting of Hong Kong must be greened to potentially cause ∼1 °C reduction in both daytime and nighttime air temperature while the same could help improve daytime pedestrian thermal comfort by at least one thermal class. We also established that higher greened facade ratio will be required to obtain similar thermal benefits in low and medium density urban settings. Also, realized benefits at pedestrians’ height can be enhanced when the vertical greening facilities are placed along podium than tower heights. Lastly, practicable urban planning recommendations were presented for the attention of urban planners and landscape architects.

Electric field driven phase transformations require two phases with a mismatch in their electric polarization, as seen in antiferroelectric-to-ferroelectric transformations, where the ferroelectric phase has a permanent polarization that is favored under field. Many other nonferroelectric dielectric materials can become electrically polarized according to their electrical susceptibility, yet such induced polarizations are not generally considered capable of enabling a phase transformation. Here we explore a susceptibility-mismatch phase transformation in a paraelectric ceramic, yttria-doped zirconia. Using in situ x-ray diffraction at $550\text{ }\ifmmode^\circ\else\textdegree\fi{}\mathrm{C}$ we show that the monoclinic-to-tetragonal transformation can be driven directly by an electric field, providing experimental evidence of a paraelectric-to-paraelectric phase transformation. Considering the $\ensuremath{\sim}1%$ mechanical strain of this transformation, the resulting electromechanical coupling may have potential for solid-state electrical actuators.

Alternative mRNA splicing is a fundamental process to increase the versatility of the genome. In humans, cardiac mRNA splicing is involved in the pathophysiology of heart failure. Mutations in the splicing factor RNA binding motif protein 20 (RBM20) cause severe forms of cardiomyopathy. To identify novel cardiomyopathy-associated splicing factors, RNA-seq and tissue-enrichment analyses were performed, which identified up-regulated expression of Sam68-Like mammalian protein 2 (SLM2) in the left ventricle of dilated cardiomyopathy (DCM) patients. In the human heart, SLM2 binds to important transcripts of sarcomere constituents, such as those encoding myosin light chain 2 (MYL2), troponin I3 (TNNI3), troponin T2 (TNNT2), tropomyosin 1/2 (TPM1/2), and titin (TTN). Mechanistically, SLM2 mediates intron retention, prevents exon exclusion, and thereby mediates alternative splicing of the mRNA regions encoding the variable proline-, glutamate-, valine-, and lysine-rich (PEVK) domain and another part of the I-band region of titin. In summary, SLM2 is a novel cardiac splicing regulator with essential functions for maintaining cardiomyocyte integrity by binding to and processing the mRNAs of essential cardiac constituents such as titin.

Consciousness is central to animal welfare concerns. Its assessment is most often conducted based on behavior, with a poor understanding of the correspondence between behavior and the neurobiological processes that underlie the subjective experience of consciousness. Recording of brain electrical activity using electrodes placed under the skull improves EEG recording by minimizing artifacts from muscular or cardiac activities, and it can now be combined with wireless recording in free-moving animals. This experiment investigated the correspondence between wireless 'under the skull' epidural EEG and the behavior of 18 five-week-old female piglets undergoing nitrous oxide (N2O) or carbon dioxide (CO2) gradual fill gas euthanasia at 25% replacement rate per minute of the chamber volume. Piglets exposed to CO2 had a peak in EEG total power ('Ptot') during the flailing stage, whereas piglets exposed to N2O had a higher EEG 95% spectral edge frequency ('F95') during their initial explorative behavior phase and a drop in EEG median frequency ('F50') after loss of posture. Loss of posture without righting attempt, as the last behavioral state observed during euthanasia, preceded the onset of transitional EEG on average by 0.9 and 3.1 min (for CO2 and N2O treatments, respectively), and the onset of isoelectric EEG by 4.5 and 6.2 min (for CO2 and N2O treatments, respectively). Paddling movements occurred shortly before and during transitional EEG but never during isoelectric EEG, whereas gasps persisted after the EEG had become isoelectric. The dynamics of EEG spectral changes were complex to interpret in relation to the degree of consciousness, but isoelectric EEG as an unequivocal indicator of unconsciousness appeared several minutes after loss of posture with no righting attempt. This leaves a window of uncertainty in regards to the potential for consciousness after loss of posture during gradual fill gas euthanasia in piglets.

There is a crucial need to identify biomarkers of epileptogenesis that will help predict later development of seizures. This work identifies two novel electrophysiological biomarkers that quantify epilepsy progression in a rat model of epileptogenesis. The long-term tetanus toxin rat model was used to show the development and remission of epilepsy over several weeks. We measured the response to periodic electrical stimulation and features of spontaneous seizure dynamics over several weeks. Both biomarkers showed dramatic changes during epileptogenesis. Electrically induced responses began to change several days before seizures began and continued to change until seizures resolved. These changes were consistent across animals and allowed development of an algorithm that could differentiate which animals would later develop epilepsy. Once seizures began, there was a progression of seizure dynamics that closely follows recent theoretical predictions, suggesting that the underlying brain state was changing over time. This research demonstrates that induced electrical responses and seizure onset dynamics are useful biomarkers to quantify dynamical changes in epileptogenesis. These tools hold promise for robust quantification of the underlying epileptogenicity and prediction of later development of seizures., This work identified two novel electrophysiological biomarkers that quantify epilepsy progression in the tetanus toxin rat model: the response to periodic electrical stimulation and features of spontaneous seizure dynamics. Both biomarkers quantify the evolution of epileptogenesis over weeks., Graphical Abstract Graphical Abstract

The exchange of radiant fluxes between different surfaces in an outdoor environment can be described with an enclosure theory. It provides a physical understanding of the relationship between different surfaces and consequent radiant fluxes within an enclosure, which is constructed by including surfaces of building facades and an imaginary surface of sky dome. Radiant fluxes on a surface can be expressed as linear combinations of view factors. This study examines the linear dependencies of urban radiant fluxes on view factors, namely Sunlit View Factor (SLVF), Green View Factor (GNVF), and Sky View Factor (SVF), using an empirical regression analysis. Results revealed a simple and linear correlation between view factors and outdoor radiant fluxes. As such, a quick estimation of the potential increase in the outdoor mean radiant temperature in relation to neighboring built environments can be achieved, and industry guidelines for designing thermally comfortable open spaces can be formulated accordi...

The transcriptome needs to be tightly regulated by mechanisms that include transcription factors, enhancers, and repressors as well as non‐coding RNAs. Besides this dynamic regulation, a large part of phenotypic variability of eukaryotes is expressed through changes in gene transcription caused by genetic variation. In this study, we evaluate genome‐wide structural genomic variants (SVs) and their association with gene expression in the human heart. We detected 3,898 individual SVs affecting all classes of gene transcripts (e.g., mRNA, miRNA, lncRNA) and regulatory genomic regions (e.g., enhancer or TFBS). In a cohort of patients (n = 50) with dilated cardiomyopathy (DCM), 80,635 non‐protein‐coding elements of the genome are deleted or duplicated by SVs, containing 3,758 long non‐coding RNAs and 1,756 protein‐coding transcripts. 65.3% of the SV‐eQTLs do not harbor a significant SNV‐eQTL, and for the regions with both classes of association, we find similar effect sizes. In case of deleted protein‐coding exons, we find downregulation of the associated transcripts, duplication events, however, do not show significant changes over all events. In summary, we are first to describe the genomic variability associated with SVs in heart failure due to DCM and dissect their impact on the transcriptome. Overall, SVs explain up to 7.5% of the variation of cardiac gene expression, underlining the importance to study human myocardial gene expression in the context of the individual genome. This has immediate implications for studies on basic mechanisms of cardiac maladaptation, biomarkers, and (gene) therapeutic studies alike.

Background: Biochemical DNA modification resembles a crucial regulatory layer among genetic information, environmental factors, and the transcriptome. To identify epigenetic susceptibility regions and novel biomarkers linked to myocardial dysfunction and heart failure, we performed the first multi-omics study in myocardial tissue and blood of patients with dilated cardiomyopathy and controls. Methods: Infinium human methylation 450 was used for high-density epigenome-wide mapping of DNA methylation in left-ventricular biopsies and whole peripheral blood of living probands. RNA deep sequencing was performed on the same samples in parallel. Whole-genome sequencing of all patients allowed exclusion of promiscuous genotype-induced methylation calls. Results: In the screening stage, we detected 59 epigenetic loci that are significantly associated with dilated cardiomyopathy (false discovery corrected P ≤0.05), with 3 of them reaching epigenome-wide significance at P ≤5×10 −8 . Twenty-seven (46%) of these loci could be replicated in independent cohorts, underlining the role of epigenetic regulation of key cardiac transcription regulators. Using a staged multi-omics study design, we link a subset of 517 epigenetic loci with dilated cardiomyopathy and cardiac gene expression. Furthermore, we identified distinct epigenetic methylation patterns that are conserved across tissues, rendering these CpGs novel epigenetic biomarkers for heart failure. Conclusions: The present study provides to our knowledge the first epigenome-wide association study in living patients with heart failure using a multi-omics approach.

Hypertrophic cardiomyopathy (HCM) is the most common genetic cardiovascular disease, which goes along with increased risk for sudden cardiac death (SCD). Despite the knowledge about the different causal genes, the relationship between individual genotypes and phenotypes is incomplete. We retrieved PubMed/Medline literatures on genotype–phenotype associations in patients with HCM and mutations in MYBPC3, MYH7, TNNT2, and TNNI3. Altogether, 51 studies with 7675 HCM patients were included in our meta-analysis. The average frequency of mutations in MYBPC3 (20%) and MYH7 (14%) was higher than TNNT2 and TNNI3 (2% each). The mean age of HCM onset for MYH7 mutation positive patients was the beginning of the fourth decade, significantly earlier than patients without sarcomeric mutations. A high male proportion was observed in TNNT2 (69%), MYBPC3 (62%) and mutation negative group (64%). Cardiac conduction disease, ventricular arrhythmia and heart transplantation (HTx) rate were higher in HCM patients with MYH7 mutations in comparison to MYBPC3 (p

Canonical transient receptor potential (TRPC) channels influence various neuronal functions. Using quantitative high‐resolution mass spectrometry, we demonstrate that TRPC1, TRPC4, and TRPC5 assemble into heteromultimers with each other, but not with other TRP family members in the mouse brain and hippocampus. In hippocampal neurons from Trpc1/Trpc4/Trpc5 ‐triple‐knockout ( Trpc1/4/5 −/− ) mice, lacking any TRPC1‐, TRPC4‐, or TRPC5‐containing channels, action potential‐triggered excitatory postsynaptic currents (EPSCs) were significantly reduced, whereas frequency, amplitude, and kinetics of quantal miniature EPSC signaling remained unchanged. Likewise, evoked postsynaptic responses in hippocampal slice recordings and transient potentiation after tetanic stimulation were decreased. In vivo , Trpc1/4/5 −/− mice displayed impaired cross‐frequency coupling in hippocampal networks and deficits in spatial working memory, while spatial reference memory was unaltered. Trpc1/4/5 −/− animals also exhibited deficiencies in adapting to a new challenge in a relearning task. Our results indicate the contribution of heteromultimeric channels from TRPC1, TRPC4, and TRPC5 subunits to the regulation of mechanisms underlying spatial working memory and flexible relearning by facilitating proper synaptic transmission in hippocampal neurons.

Ceria doped zirconia has been shown to exhibit enhanced shape memory properties in small volume structures such as particles and micropillars. Here those properties are translated into macroscopic materials through the fabrication of zirconia foams by ice templating. Directional freezing is used to produce foams with micron-sized pores and struts that can locally take advantage of the shape memory effect due to their fine scale. The foams are subjected to thermal cycling and x-ray diffraction analysis to evaluate the martensitic transformation that underlies shape memory properties, and are found to survive the transformation through multiple cycles. Keywords: Freeze casting; Foams; Shape memory effect; Martensitic phase transformation

High-density urban environment affects urban microclimate, and thermal comfort of outdoor spaces. Shading by urban structure is the primary measure to reduce daytime mean radiant temperature (MRT) by blocking direct solar radiation. However, it is believed that overly-restricted Sky View Factor (SVF) limits the radiative cooling by the sky of lower effective temperature. Therefore, this study aimed at examining the relationship between SVF and MRT in built environment. To investigate the dependence of MRT on SVF via radiant fluxes, this study performed field measurement of six directional long-wave, short-wave fluxes and MRT across a number of outdoor spaces under shaded in densely built environment. Regression analysis was employed in examining desired relationships. Across different open spaces, MRT would increase by 1.6 K per 10 W/m2 increase of either long-wave or short-wave radiant fluxes. Global solar radiation and air temperature are the most influential meteorological parameters affecting radiant fluxes components in MRT. Besides, this study suggests the use of long-wave mean radiant temperature (LMRT) representing effective surface temperature of surrounding objects. LMRT generally follows the temporal pattern of reference air temperature. In a case study, if SVF is decreased by 0.1, the weighted sum of long-wave fluxes would be increased by 10 W/m2. This would bring an increase of 1.6 K in MRT. But, SVF has weak explanatory power to variations in short-wave fluxes when under shaded. Larger SVF is preferred for cooling open space in hot and humid region if direct sunlight is already blocked by urban morphology.

To investigate the impact of sleep onset and offset on the rate of epileptiform discharges (ED) in idiopathic generalized epilepsies (IGE).We studied the temporal distribution of EDs with mixed-effects Poisson regression modeling in a cohort of patients diagnosed with IGE who underwent 24-hour ambulatory electroencephalography (EEG) recordings. We defined the mean number discharges per hour per subject as the mean ED rate. The association between each hour and the mean ED rate was quantified with incidence rate ratio (IRR) as the metric. We calculated the IRR of each hourly block for the total cohort in relation to sleep onset and offset. Finally, we admitted secondary risk factors into our Poisson regression model and quantified changes in IRR in order to investigate the impact of those variables on the outcome. The secondary risk factors included: epilepsy syndrome, duration of seizure freedom, duration of epilepsy, number of antiepileptic drugs (AED), type of AED, and age.A total of 39 patients with a mean age of 29.1 y (SD = 10.1) were studied. The distribution of ED rate demonstrated a highly significant abrupt increase in the first hour after sleep onset (IRR = 3.96; p 0.001). On the contrary, the ED rate significantly dropped in the second hour after the sleep offset compared with the last hour block before sleep offset (IRR = 0.39; p 0.001). None of the secondary risk factors demonstrated any significant impact on this pattern.Sleep onset is a very significant trigger for the generation of EDs in IGE.Our results support the hypothesis that there is a "critical zone of vigilance" in the sleep-wake boundary from which generalized EDs are more likely to emerge.

Small volume samples of zirconia can survive stress-induced martensitic transformation without cracking, which enables in-depth explorations of martensite mechanics using micro-scale specimens. Here we present a systematic investigation of the orientation dependence of tetragonal crystals undergoing a uniaxial stress-driven martensitic transformation to the monoclinic phase, in single crystal zirconia pillars doped with yttria and titania. The Young’s modulus, martensitic transformation stress and transformation strain are highly dependent on the crystallographic orientation, and generally align with expectations based on known tensor properties and transformation crystallography. However, in some orientations, fracture or plastic slip are apparently preferred to martensitic transformation, and thus crystallography favors certain orientations if superelasticity or shape memory properties are specifically desired in zirconia ceramics. Accepted Version

Routine genetic testing in Dilated Cardiomyopathy (DCM) has recently become reality using Next-Generation Sequencing. Several studies have explored the relationship between genotypes and clinical phenotypes to support risk estimation and therapeutic decisions, however, most studies are small or restricted to a few genes. This study provides to our knowledge the first systematic meta-analysis on genotype-phenotype associations in DCM. We retrieved PubMed/Medline literature on genotype–phenotype associations in patients with DCM and mutations in LMNA, PLN, RBM20, MYBPC3, MYH7, TNNT2 and TNNI3. We summarized and extensively reviewed all studies that passed selection criteria and performed a meta-analysis on key phenotypic parameters. Together, 48 studies with 8097 patients were included. Furthermore, we reviewed recent studies investigating genotype-phenotype associations in DCM patients with TTN mutations. The average frequency of mutations in the investigated genes was between 1 and 5 %. The mean age of DCM onset was the beginning of the fifth decade for all genes. Heart transplantation (HTx) rate was highest in LMNA mutation carriers (27 %), while RBM20 mutation carriers were transplanted at a markedly younger age (mean 28.5 years). While 73 % of DCM patients with LMNA mutations showed cardiac conduction diseases, low voltage was the reported ECG hallmark in PLN mutation carriers. The frequency of ventricular arrhythmia in DCM patients with LMNA (50 %) and PLN (43 %) mutations was significantly higher. The penetrance of DCM phenotype in subjects with TTN truncating variants increased with age and reached 100 % by age of 70. A pooled analysis of available genotype-phenotype data shows a higher prevalence of sudden cardiac death (SCD), cardiac transplantation, or ventricular arrhythmias in LMNA and PLN mutation carriers compared to sarcomeric gene mutations. This study will further support the clinical interpretation of genetic findings.

Next-generation sequencing (NGS) is getting routinely used in the diagnosis of hereditary diseases, such as human cardiomyopathies. Hence, it is of utter importance to secure high quality sequencing data, enabling the identification of disease-relevant mutations or the conclusion of negative test results. During the process of sample preparation, each protocol for target enrichment library preparation has its own requirements for quality control (QC); however, there is little evidence on the actual impact of these guidelines on resulting data quality. In this study, we analyzed the impact of QC during the diverse library preparation steps of Agilent SureSelect XT target enrichment and Illumina sequencing. We quantified the parameters for a cohort of around 600 samples, which include starting amount of DNA, amount of sheared DNA, smallest and largest fragment size of the starting DNA; amount of DNA after the pre-PCR, and smallest and largest fragment size of the resulting DNA; as well as the amount of the final library, the corresponding smallest and largest fragment size, and the number of detected variants. Intriguingly, there is a high tolerance for variations in all QC steps, meaning that within the boundaries proposed in the current study, a considerable variance at each step of QC can be well tolerated without compromising NGS quality.