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Muscle contraction is driven by the molecular machinery of the sarcomere. As phosphorylation is a critical regulator of muscle function, the identification of regulatory kinases is important for understanding sarcomere biology. Pathogenic variants in alpha kinase 3 (ALPK3) cause cardiomyopathy and musculoskeletal disease, but little is known about this atypical kinase. Here we show that ALPK3 is an essential component of the M-band of the sarcomere and define the ALPK3-dependent phosphoproteome. ALPK3 deficiency impaired contractility both in human cardiac organoids and in the hearts of mice harboring a pathogenic truncating Alpk3 variant. ALPK3-dependent phosphopeptides were enriched for sarcomeric components of the M-band and the ubiquitin-binding protein sequestosome-1 (SQSTM1) (also known as p62). Analysis of the ALPK3 interactome confirmed binding to M-band proteins including SQSTM1. In human pluripotent stem cell-derived cardiomyocytes modeling cardiomyopathic ALPK3 mutations, sarcomeric organization and M-band localization of SQSTM1 were abnormal suggesting that this mechanism may underly disease pathogenesis.

Glioblastoma (GBM) brain tumors contain a subpopulation of self-renewing multipotent Glioblastoma stem-like cells (GSCs) that are believed to drive the near inevitable recurrence of GBM. We previously engineered temperature responsive scaffolds based on the polymer poly(N-isopropylacrylamide-co-Jeffamine M-1000 acrylamide) (PNJ) for the purpose of enriching GSCs in vitro from patient-derived samples. Here, we used PNJ scaffolds to study microenvironmental regulation of self-renewal and radiation response in patient-derived GSCs representing classical and proneural subtypes. GSC self-renewal was regulated by the composition of PNJ scaffolds and varied with cell type. PNJ scaffolds protected against radiation-induced cell death, particularly in conditions that also promoted GSC self-renewal. Additionally, cells cultured in PNJ scaffolds exhibited increased expression of the transcription factor HIF2α, which was not observed in neurosphere culture, providing a potential mechanistic basis for differences in radio-resistance. Differences in PNJ regulation of HIF2α in irradiated and untreated conditions also offered evidence of stem plasticity. These data show PNJ scaffolds provide a unique biomaterial for evaluating dynamic microenvironmental regulation of GSC self-renewal, radioresistance, and stem plasticity.

Schizophrenia has a multifactorial etiology, involving a polygenic architecture. The potential benefit of whole genome sequencing (WGS) in schizophrenia and other psychotic disorders is not well studied. We investigated the yield of clinical WGS analysis in 251 families with a proband diagnosed with schizophrenia (N = 190), schizoaffective disorder (N = 49), or other conditions involving psychosis (N = 48). Participants were recruited in Israel and USA, mainly of Jewish, Arab, and other European ancestries. Trio (parents and proband) WGS was performed for 228 families (90.8%); in the other families, WGS included parents and at least two affected siblings. In the secondary analyses, we evaluated the contribution of rare variant enrichment in particular gene sets, and calculated polygenic risk score (PRS) for schizophrenia. For the primary outcome, diagnostic rate was 6.4%; we found clinically significant, single nucleotide variants (SNVs) or small insertions or deletions (indels) in 14 probands (5.6%), and copy number variants (CNVs) in 2 (0.8%). Significant enrichment of rare loss-of-function variants was observed in a gene set of top schizophrenia candidate genes in affected individuals, compared with population controls (N = 6,840). The PRS for schizophrenia was significantly increased in the affected individuals group, compared to their unaffected relatives. Last, we were also able to provide pharmacogenomics information based on CYP2D6 genotype data for most participants, and determine their antipsychotic metabolizer status. In conclusion, our findings suggest that WGS may have a role in the setting of both research and genetic counseling for individuals with schizophrenia and other psychotic disorders and their families.

Com muita honra, apresento a Profa. Fernanda nesta entrevista, minha amiga de longa data, com quem tive a grata satisfação de trabalhar nos cursos de especialização em Ortodontia da Universidade de Marília (Marília/SP) e da Ponti- fícia Universidade Católica Madre y Maestra, em Santiago de Los Caballeros (República Dominicana). As contribuições da Profa. Fernanda para a Ortodontia são maiores do que a sua estatura, em razão da sua dedicação ao ensino e à pesquisa. Um bom exemplo das suas contribuições é a classificação dos estágios de maturação da sutura palatina mediana, proposta em 2013 por ela e colaboradores. Essa classificação teve, e vem tendo, grande aceitação por acadêmicos, clínicos e pesquisadores das diferentes partes do mundo.

Faster, and simpler methods that determine the susceptibility of bacterial pathogens to antibiotics are needed to enable more effective treatment of bacterial infections and reduce the overuse of antibiotics. Here, we demonstrate a simple method for rapidly detecting bacteria and simultaneously determining their antibiotic susceptibility status. We demonstrate the method withE. coli, a high-impact pathogen, and meropenem, a member of the carbapenem class of antibiotics which is usually reserved for multi-drug resistant infections. Inclusion of a fluorogenic oligonucleotide substrate for endonuclease I, a well-conserved nuclease of the Enterobacteriaceae family, in a tryptic soy agar-based media enabled fluorescence-based detection ofE. coligrowth within 9 hours. Plating established carbapenem-resistant and -susceptibleE. colistrains on this media with and without inclusion of meropenem, followed by fluorescent imaging yielded a clear phenotypic measure of the antibiotic susceptibility status of the strains in 9 hours. In addition to its simplicity and fast turnaround time, advantages of this approach include its ability to provide a measure of the bacterial load of the tested samples, and its easy integration into current microbiology laboratory workflows.

In recent years, histone deacetylase inhibitors (HDACi) have garnered considerable interest for the treatment of adult and pediatric malignant brain tumors. However, owing to their broad-spectrum nature and inability to effectively penetrate the blood-brain barrier, HDACi have failed to provide significant clinical benefit to glioblastoma (GBM) patients to date. Moreover, global inhibition of HDACs results in widespread toxicity, highlighting the need for selective isoform targeting. While no isoform-specific HDACi are currently available, the second-generation hydroxamic acid-based HDACi quisinostat possesses sub-nanomolar specificity for class I HDAC isoforms, particularly HDAC1 and 2. Recently, we demonstrated that HDAC1 is the essential HDAC in GBM. Here, we provide the first report on the neuro-pharmacokinetic, pharmacodynamic and radiation-sensitizing properties of quisinostat in preclinical models of GBM. We demonstrate that quisinostat is a well-tolerated and brain-penetrant molecule that significantly extends survival when administered in combination with radiationin vivo. The pharmacokinetic-pharmacodynamic-efficacy relationship was established by correlating free drug concentrations and evidence of target modulation in the brain with survival benefit. Together, these data provide a strong rationale for clinical development of quisinostat as a radiosensitizer for the treatment of GBM.

SUMMARYPathogenic variants in alpha kinase 3 (ALPK3) cause cardiomyopathy and musculoskeletal disease. How ALPK3 mutations result in disease remains unclear because little is known about this atypical kinase. Using a suite of engineered human pluripotent stem cells (hPSCs) we show that ALPK3 localizes to the M-Band of the sarcomere. ALPK3 deficiency disrupted sarcomeric organization and calcium kinetics in hPSC-derived cardiomyocytes and reduced force generation in cardiac organoids. Phosphoproteomic profiling identified ALPK3-dependant phospho-peptides that were enriched for sarcomeric components of the M-band and the ubiquitin-binding protein SQSTM1. Analysis of the ALPK3 interactome confirmed binding to M-band proteins including SQSTM1. Importantly, in hPSC-derived cardiomyocytes modeling ALPK3 deficiency and cardiomyopathic ALPK3 mutations, sarcomeric organization and M-band localization ofSQSTM1 were abnormal. These data suggest ALPK3 has an integral role in maintaining sarcomere integrity and proteostasis in striated muscle. We propose this mechanism may underly disease pathogenesis in patients with ALPK3 variants.

INTRODUCTION The purpose of the present study was to evaluate the long-term variations in maxillary second molar position in untreated subjects with normal occlusion. SETTING AND SAMPLE POPULATION A sample of 39 subjects (18 females and 21 males) selected from the University of Michigan Growth Study (UMGS) was followed longitudinally with digital dental casts at 3 observation times: T1, when the maxillary permanent second molars were fully erupted, T2, last observation available in the longitudinal series (38 subjects), and T3, at least 20 years after T2 (12 subjects). MATERIALS AND METHODS Digital measurements were recorded with an open-source software. Outcome variables were sagittal and transverse inclinations of the upper second molars. Two mixed-effect models were performed. RESULTS The maxillary second molars had a distolingual inclination at T1, T2 and T3. Sagittal and transverse inclination showed progressive significant uprighting from T1 through T3 (P

International audience; Phosphorylation of cardiac myosin binding protein-C (cMyBP-C) regulates cardiac contraction through modulation of actomyosin interactions mediated by the protein's amino terminal (N′)-region (C0-C2 domains, 358 amino acids). On the other hand, dephosphorylation of cMyBP-C during myocardial injury results in cleavage of the 271 amino acid C0-C1f region and subsequent contractile dysfunction. Yet, our current understanding of amino terminus region of cMyBP-C in the context of regulating thin and thick filament interactions is limited. A novel cardiac-specific transgenic mouse model expressing cMyBP-C, but lacking its C0-C1f region (cMyBP-C∆C0-C1f), displayed dilated cardiomyopathy, underscoring the importance of the N′-region in cMyBP-C. Further exploring the molecular basis for this cardiomyopathy, in vitro studies revealed increased interfilament lattice spacing and rate of tension redevelopment, as well as faster actin-filament sliding velocity within the C-zone of the transgenic sarcomere. Moreover, phosphorylation of the unablated phosphoregulatory sites was increased, likely contributing to normal sarcomere morphology and myoarchitecture. These results led us to hypothesize that restoration of the N′-region of cMyBP-C would return actomyosin interaction to its steady state. Accordingly, we administered recombinant C0-C2 (rC0-C2) to permeabilized cardiomyocytes from transgenic, cMyBP-C null, and human heart failure biopsies, and we found that normal regulation of actomyosin interaction and contractility was restored. Overall, these data provide a unique picture of selective perturbations of the cardiac sarcomere that either lead to injury or adaptation to injury in the myocardium.

This is a summary report of clinical and regulatory issues discussed at the 2018 NINDS workshop, entitled “Accelerating Therapies for Antiepileptogenesis and Disease Modification.” The intent of the workshop was to optimize and accelerate development of therapies for antiepileptogenesis (AEG) and disease modification in the epilepsies. The working group discussed nomenclature for antiepileptogenic therapies, subdividing them into “antiepileptogenic therapies” and “disease modifying therapies,” both of which are urgently needed. We use the example of traumatic brain injury to explain issues and complexities in designing a trial for disease‐preventing antiepileptogenic therapies, including identifying timing of intervention, selecting the appropriate dose, and the need for biomarkers. We discuss the recent trials of vigabatrin to prevent onset and modify epilepsy outcome in children with tuberous sclerosis (Epistop and PreVeNT). We describe a potential approach to a disease modification trial in adults, using patients with temporal lobe epilepsy. Finally, we discuss regulatory hurdles for antiepileptogenesis and disease‐modifying trials.

BACKGROUND: Neonatal mouse cardiomyocytes undergo a metabolic switch from glycolysis to oxidative phosphorylation, which results in a significant increase in reactive oxygen species (ROS) production that induces DNA damage. These cellular changes contribute to cardiomyocyte cell cycle exit and loss of the capacity for cardiac regeneration. The mechanisms that regulate this metabolic switch and the increase in ROS production have been relatively unexplored. Current evidence suggests that elevated ROS production in ischemic tissues occurs due to accumulation of the mitochondrial metabolite succinate during ischemia via succinate dehydrogenase (SDH), and this succinate is rapidly oxidized at reperfusion. Interestingly, mutations in SDH in familial cancer syndromes have been demonstrated to promote a metabolic shift into glycolytic metabolism, suggesting a potential role for SDH in regulating cellular metabolism. Whether succinate and SDH regulate cardiomyocyte cell cycle activity and the cardiac metabolic state remains unclear. METHODS: Here, we investigated the role of succinate and succinate dehydrogenase (SDH) inhibition in regulation of postnatal cardiomyocyte cell cycle activity and heart regeneration. RESULTS: Our results demonstrate that injection of succinate in neonatal mice results in inhibition of cardiomyocyte proliferation and regeneration. Our evidence also shows that inhibition of SDH by malonate treatment after birth extends the window of cardiomyocyte proliferation and regeneration in juvenile mice. Remarkably, extending malonate treatment to the adult mouse heart following myocardial infarction injury results in a robust regenerative response within 4 weeks following injury via promoting adult cardiomyocyte proliferation and revascularization. Our metabolite analysis following SDH inhibition by malonate induces dynamic changes in adult cardiac metabolism. CONCLUSIONS: Inhibition of SDH by malonate promotes adult cardiomyocyte proliferation, revascularization, and heart regeneration via metabolic reprogramming. These findings support a potentially important new therapeutic approach for human heart failure.

To compare the transverse dental and skeletal changes in patients treated with bone-anchored palatal expander (bone-borne, BB) compared to patients treated with tooth and bone-anchored palatal expanders (tooth-bone-borne, TBB) using cone-beam computer tomography (CBCT) and 3D image analysis.The sample comprised 30 patients with transverse maxillary discrepancy treated with two different types of appliances: bone-borne (Group BB) and tooth-bone-borne (Group TBB) expanders. CBCT scans were acquired before (T1) and after completion of maxillary expansion (T2); the interval was 5.4 ± 3.4 and 6.2 ± 2.1 months between the T1 and the T2 scans of Group TBB (tooth-bone-borne) and Group BB (bone-borne), respectively. Transverse, anteroposterior and vertical linear and angular three-dimensional dentoskeletal changes were assessed after cranial base superimposition.Both groups displayed marked transverse skeletal expansion with a greater ratio of skeletal to dental changes. Greater changes at the nasal cavity, zygoma and orbital levels were found in Group BB. A relatively parallel sutural opening in an anterior-posterior direction was observed in Group TBB; however, the Group BB presented a somewhat triangular (V-shaped) opening of the suture that was wider anteriorly. Small downward-forward displacements were observed in both groups. Asymmetric expansion occurred in approximately 50% of the patients in both groups.Greater skeletal vs dental expansion ratio and expansion of the circummaxillary regions were found in Group BB, the group in which a bone-borne expander was used. Both groups presented skeletal and dental changes, with a similar amount of posterior palate expansion. Asymmetric expansion was observed in both groups.

Objectives To determine dentoalveolar and skeletal effects produced in mature patients by the Carriere Motion Class III (CM3) appliance followed by fixed appliances. Materials and Methods This retrospective study evaluated 32 patients at three time points: T1 (initial), T2 (removal of CM3), and T3 (posttreatment). Serial cephalograms were traced and digitized, and best-fit regional superimpositions were constructed. Eleven linear and 7 angular variables were measured. The starting forms of the CM3 patients were compared with a sample of untreated subjects with normal occlusions and well-balanced faces. Results The CM3 phase lasted 6.3 months, followed by a phase of fixed appliances lasting 12.9 months; the total duration of treatment was 19.2 months. Minimal skeletal changes were measured sagittally, with only a slight increase in lower anterior facial height observed during treatment. Most treatment changes were dentoalveolar in nature. Wits appraisal increased 4.0 mm during treatment. The molar relationship improved by 6.0 mm during phase I, a value that rebounded slightly during phase II, resulting in an improvement toward Class I of 4.8 mm. Best-fit regional superimpositions revealed anterior movement of upper molars relative to the maxilla and posterior movement of lower molars relative to the mandible. Conclusions The Carriere Motion Class III appliance is an effective and efficient method of resolving occlusal problems in minimally growing Class III patients. Primary treatment effects are dentoalveolar in nature with minimal skeletal alterations.

Objective The aim of this study was to develop a prediction model that combines the information derived from chronological age (analysed as a curvilinear variable), gender and the CVM method to predict mandibular growth. Settings and sample population 50 participants (29 females, 21 males) were selected from the AAOF Craniofacial Growth Legacy Collection, the Michigan Growth Study and the Denver Child Growth study. Materials and methods In this investigation, 456 lateral cephalograms were analysed by applying a mixed effect model. The outcome variable was the annualized increment in total mandibular length (Co-Gn) during the year following the lateral cephalogram on which the cervical stage and chronological age were evaluated. The predictive variables were chronological age up to the fifth order, gender, stage of cervical vertebral maturation, as well as interactions between age and gender, age and cervical stage, and gender and cervical stage. Results Cervical stage, chronological age up to the fourth order, gender, and the interaction between age and gender were significant predictors of annualized increments in mandibular length. The annualized increment in Co-Gn was significantly greater for CS 3 when compared to all other cervical stages. Further, annualized increments in Co-Gn for CS 1 and CS 2 were significantly greater when compared to CS 5. Conclusions Cervical stage, chronological age and gender can be used jointly to predict the annualized increment in mandibular growth. Cervical stage 3 exhibited the greatest annualized increase in mandibular length.

Introduction During the decision-making process, physicians rely on heuristics that consist of simple, useful procedures for solving problems, intuitive shortcuts that produce reliable decisions based on limited information. In clinical situations characterized by a high degree of uncertainty such as those encountered in orthodontics, cognitive biases and judgment errors related to heuristics are not uncommon. This study aimed at promoting trust in the effective interface between the intuitive reasoning of the orthodontic practitioner and the computational heuristics emerging from simple statistical models. Methods We propose an integrative model based on the interaction between clinical reasoning and 2 computational tools, cluster analysis and fast-and-frugal trees, to extract a structured craniofacial representation of untreated subjects with Class III malocclusion and to forecast the worsening of the malocclusion over time. Results Cluster analysis of cephalometric values from 144 growing subjects with Class III malocclusion followed longitudinally (T1: mean age, 10.2 ± 1.9 years; T2: mean age, 13.8 ± 2.7 years) produced 3 morphologic subgroups with predominant sagittal, vertical, and slight maxillomandibular imbalances. Fast-and-frugal trees applied to different subgroups extracted heuristics that improved the prediction of key features associated with adverse craniofacial growth. Conclusions Provided that cephalometric values are placed in the appropriate framework, the matching between simple and fast computational approaches and clinical reasoning could help the intuitive logic, perception, and cognitive inferences of orthodontic practitioners on the outcome of patients affected by Class III disharmony, decreasing errors associated with flawed judgments and improving the accuracy of decision making.

Aim The aim of this study was to analyze dental and soft tissue profile development in children with normal occlusions to establish age- and gender-specific reference intervals for German children during their active growth period. Subjects and methods The study group consisted of a sample of 31 untreated Caucasian subjects with normal occlusions. Dental casts were analyzed at four different stages of dentitional development. Extraoral profile photographs were available for 19 subjects at stages T2–T4. In these subjects 11 angular measurements and 14 indices were analyzed. Statistical comparisons of gender-specific differences were performed by Mann–Whitney U tests (p ≤ 0.05). Results Upper and lower posterior and total arch perimeters were recorded to be significantly larger in male subjects until the late mixed dentition. Subsequently, there was a tendency toward larger dimensions in males for those parameters. Upper and lower intercanine, interpremolar and intermolar widths were significantly larger in males throughout the entire observation period. There were no statistically significant gender differences with regard to most angular measurements in the dental arches, including molar rotation, palatal volume, overbite, overjet and molar relationship at later dental stages. Conclusion In untreated subjects with normal occlusion, dental arch and soft tissue parameters can be considered age-dependent. For some dental parameters, gender-specific differences were found that should be taken into consideration during diagnosis and treatment planning of growing children. The obtained longitudinal data of untreated children provide useful information for orthodontic diagnosis, treatment planning and future research projects.

Methods that enable rapid detection of SARS-CoV-2 provide valuable tools for detecting and controlling Covid-19 outbreaks and also facilitate more effective treatment of infected individuals. The predominant approaches developed use PCR to detect viral nucleic acids or immunoassays to detect viral proteins. Each approach has distinct advantages and disadvantages, but alternatives that do not share the same limitations could enable substantial improvements in outbreak detection and management. For instance, methods that have comparable sensitivity to PCR, but that are not prone to the false-positive results that stem from the tendency of PCR to detect molecular degradation products could improve accurate identification of infected individuals. An alternative approach with potential to achieve this entails harnessing the unique enzymatic properties of SARS-CoV-2 enzymes to generate SARS-Cov-2-specific signals that indicate the presence of the virus. This route benefits from the high sensitivity provided by enzymatic signal amplification and also the fact that signal is generated only by intact viral enzymes, not degradation products. Here, we demonstrate enzymatic reaction conditions that enable the preferential detection of NendoU of SARS-CoV-2, versus several of its orthologues, with a fluorogenic oligonucleotide substrate. These compositions provide a possible technical foundation for a novel approach for detecting SARS-CoV-2 that has distinct advantages from current approaches.

OBJECTIVES: To determine the skeletal and dental changes with Microimplant Assisted Rapid Palatal Expansion (MARPE) appliances in growing and non-growing patients using cone beam computed tomography (CBCT) and 3D imaging analysis. METHODS: The sample was comprised of 25 patients with transverse maxillary discrepancy treated with Maxillary Skeletal Expander (MSE, a type of MARPE appliance). CBCT scans were taken before and after maxillary expansion; the interval was 6 ± 4.3 months. The sample was divided into growing and non-growing groups using cervical vertebrae and midpalatal suture maturation. Linear and angular three dimensional dentoskeletal changes were assessed after cranial base superimposition. The treatment groups were compared with Independent Sample t-test (P < .05). RESULTS: Both groups displayed marked transverse changes with a similar ratio of skeletal to dental transverse changes and parallel sutural opening from the PNSANS; a similar amount of expansion occurred in the anterior and the posterior regions of the maxilla. The maxilla expanded skeletally without rotational displacements in both groups. The small downward-forward displacements were similar in both groups, except that the growing group had a significantly greater vertical displacement of the canines (1.7 ±1.0 mm in growing patients, 0.6 ±0.8 mm in non-growing patients; P = 0.02) and ANS (1.1 ±0.6 mm in the growing group, 0.5 ±0.5 mm in the non-growing group; P = 0.004). CONCLUSIONS: Treatment of patients with MARPE appliance is effective in growing and non-growing patients. While greater skeletal and dental changes were observed in growing patients, a similar ratio of skeletal to dental transverse changes was observed in both groups.

Soil climate, as quantified by soil temperature (TS) and water content (θ), exerts important controls on critical zone processes. It may be sensitive to variations in local slope and aspect (SA), but this attribute remains poorly quantified at the local scale and unresolved in large‐scale models. Estimation of SA effects on soil climate across multiple scales may facilitated using topographically modified, incoming clear‐sky solar radiation (SR,CS,T). We established six paired automated soil climate monitoring stations on opposing north‐facing (NF) and south‐facing (SF) slopes (4 yr) and collected spatial TS and θ data within the hectare surrounding four stations (2 yr) to measure SA effects on soil climate. Results were compared with physically based simulations and evaluated in the context of SR,CS,T. Spatial θ data were more variable than Ts, and both were consistent with values from continuous monitoring stations. On average, the SF TS was much greater (4.7 °C) and the annual summer drought longer (36 d) than on the adjacent NF aspect. Seasonal variations of TS and θ were different from each other and also different from SR,CS,T. Local conditions, including snow cover, precipitation patterns, and soil properties, largely controlled seasonal variations of TS and θ, which cannot be predicted from SR,CS,T. This indicates that realistic simulation of many critical zone processes requires high‐resolution inputs. Simulations captured first‐order SA effects and could be useful for estimating SA effects in lieu of field monitoring.

Insight into the cellular and circuit mechanisms underlying development of temporal lobe epilepsy (TLE) will provide a foundation for improved therapies. We studied a model in which an episode of prolonged seizures is followed by recovery lasting two weeks before emergence of spontaneous recurrent seizures. We focused on the interval between the prolonged seizures and the late onset recurrent seizures. We investigated the hippocampal mossy fiber CA3 pyramidal cell microcircuit in models spanningin vitro,in vivo, andex vivopreparations. Expression of channelrhodopsin-2 in the dentate granule cells ofDGC ChRmice enabled the selective activation of mossy fiber axons.In vivostudies revealed marked potentiation of mossy fiber evoked field potentials in hippocampal CA3 beginning within hours following seizures, a potentiation which persisted at least 7 d. Stimulation of mossy fibers in hippocampal slicesin vitrousing patterns of activity mimicking seizures induced LTP not only of the monosynaptic EPSC but also of the disynaptic IPSC of CA3 pyramidal cells.Ex vivostudies of slices isolated following seizures revealed evidence of LTP of mossy fiber evoked EPSC and disynaptic IPSC of CA3 pyramidal cells. We suggest that activation of dentate granule cells during seizures induces these plasticitiesin vivoand the retained balance of synaptic excitation and inhibition limits excessive activation of CA3 pyramidal cells, thereby protecting animals from spontaneous recurrent seizures at this interval following status epilepticus.

Introduction This study aimed to determine the volumetric effects on the upper airways of growing patients with Class II malocclusion treated with the Herbst appliance (HA). Methods Volumetric measurements of the upper airways of 42 skeletal Class II malocclusion patients (mean age: 13.8 ± 1.2 years; ranging from 12.0 to 16.9 years) were assessed using cone-beam computed tomography scans acquired before treatment (T0) and approximately 1 year later (T1). The sample comprised a Herbst appliance group (HA group [HAG]; n = 24), and a comparison group (comparison group [CG]; n = 18) of orthodontic patients who had received dental treatments other than mandibular advancement with dentofacial orthopedics. Results In CG, nasopharynx and oropharynx volumes decreased slightly during the observation period (9% and 3%, respectively), whereas the nasal cavity volume increased significantly (12%; P = 0.046). In HAG, there was an increase in the volume of all regions (nasal cavity, 5.5%; nasopharynx, 11.7%; and oropharynx, 29.7%). However, only the oropharynx showed a statistically significant increase (P = 0.003), presenting significant volumetric changes along the time (T1-T0) in HAG. Conclusion Mandibular advancement with the HA significantly increased the volume of the oropharynx, but no significant volumetric modifications were observed in the nasal cavity and nasopharynx.

Visual Abstract, Highlights • Extensive LA fibrosis is related to an increase in LA size and decrease in total LA EF. This supports the contention that the LA is not merely a bystander of MR but rather is actively involved in the pathophysiology of MR. • Chymase is plentiful in the human heart, especially in the LA, and correlates with extracellular matrix accumulation in primary MR. • Because of the unreliability of the LV ejection fraction in primary MR, future studies may consider LA size and total LA EF for timing of surgical intervention of asymptomatic moderate to severe MR., Summary Increasing left atrial (LA) size predicts outcomes in patients with isolated mitral regurgitation (MR). Chymase is plentiful in the human heart and affects extracellular matrix remodeling. Chymase activation correlates to LA fibrosis, LA enlargement, and a decreased total LA emptying fraction in addition to having a potential intracellular role in mediating myofibrillar breakdown in LA myocytes. Because of the unreliability of the left ventricular ejection fraction in predicting outcomes in MR, LA size and the total LA emptying fraction may be more suitable indicators for timing of surgical intervention.

Objectives: To determine the treatment effects produced in Class II patients by the Carriere® Motion 3D™ appliance (CMA) followed by full fixed appliances (FFA). Materials and Methods: This retrospective study evaluated 34 adolescents at three time points: T1 (pretreatment), T2 (removal of CMA), and T3 (posttreatment). The comparison group comprised 22 untreated Class II subjects analyzed at T1 and T3. Serial cephalograms were traced and digitized, and 12 skeletal and 6 dentoalveolar measures were compared. Results: Phase I with CMA lasted 5.2 ± 2.8 months; phase II with FFA lasted 13.0 ± 4.2 months. CMA treatment restricted the forward movement of the maxilla at point A. There was minimal effect on the sagittal position of the chin at pogonion. The Wits appraisal improved toward Class I by 2.1 mm during the CMA phase but not during FFA. Lower anterior facial height increased twice as much in the treatment group as in controls. A clockwise rotation (3.9°) of the functional occlusal plane in the treatment group occurred during phase I; a substantial rebound (−3.6°) occurred during phase II. Overjet and overbite improved during treatment, as did molar relationship; the lower incisors proclined (4.2°). Conclusions: The CMA appliance is an efficient and effective way of correcting Class II malocclusion. The changes were mainly dentoalveolar in nature, but some skeletal changes also occurred, particularly in the sagittal position of the maxilla and in the vertical dimension.

Heart failure represents the end result of different pathophysiologic processes, which culminate in functional impairment. Regardless of its aetiology, the presentation of heart failure usually involves symptoms of pump failure and congestion, which forms the basis for clinical diagnosis. Pathophysiologic descriptions of heart failure with reduced ejection fraction (HFrEF) are being established. Most commonly, HFrEF is centred on a reactive model where a significant initial insult leads to reduced cardiac output, further triggering a cascade of maladaptive processes. Predisposing factors include myocardial injury of any cause, chronically abnormal loading due to hypertension, valvular disease, or tachyarrhythmias. The pathophysiologic processes behind remodelling in heart failure are complex and reflect systemic neurohormonal activation, peripheral vascular effects and localised changes affecting the cardiac substrate. These abnormalities have been the subject of intense research. Much of the translational successes in HFrEF have come from targeting neurohormonal responses to reduced cardiac output, with blockade of the renin-angiotensin-aldosterone system (RAAS) and beta-adrenergic blockade being particularly fruitful. However, mortality and morbidity associated with heart failure remains high. Although systemic neurohormonal blockade slows disease progression, localised ventricular remodelling still adversely affects contractile function. Novel therapy targeted at improving cardiac contractile mechanics in HFrEF hold the promise of alleviating heart failure at its source, yet so far none has found success. Nevertheless, there are increasing calls for a proximal, 'cardiocentric' approach to therapy. In this review, we examine HFrEF therapy aimed at improving cardiac function with a focus on recent trials and emerging targets.

The non-contractile cytoskeleton in cardiomyocytes is comprised of cytoplasmic actin, microtubules, and intermediate filaments. In addition to providing mechanical support to these cells, these structures are important effectors of tension-sensing and signal transduction and also provide networks for the transport of proteins and organelles. The majority of our knowledge on the function and structure of these cytoskeletal networks comes from research on proliferative cell types. However, in recent years, researchers have begun to show that there are important cardiomyocyte-specific functions of the cytoskeleton. Here we will discuss the current state of cytoskeletal biology in cardiomyocytes, as well as research from other cell types, that together suggest there is a wealth of knowledge on cardiac health and disease waiting to be uncovered through exploration of the complex signaling networks of cardiomyocyte non-sarcomeric cytoskeletal proteins.

Integrated watershed models can be used to calculate streamflow generation in snow‐dominated mountainous catchments. Parameterization of water flow is often complicated by the lack of information on subsurface hydraulic properties. In this study, bulk density optimization was used to determine hydraulic parameters for the upper and lower regolith in the GEOtop model. The methodology was tested in two small catchments in the Dry Creek Watershed in Idaho and the Libby Creek Watershed in Wyoming. Modelling efficiencies for profile‐average soil–water content for the two catchments were between 0.52 and 0.64. Modelling efficiencies for stream discharge (cumulative stream discharge) were 0.45 (0.91) and 0.54 (0.94) for the Idaho and Wyoming catchments, respectively. The calculated hydraulic properties suggest that lateral flow across the upper–lower regolith interface is an important driver of streamflow in both the Idaho and Wyoming watersheds. The overall calibration procedure is computationally efficient because only two bulk density values are optimized. The two‐parameter calibration procedure was complicated by uncertainty in hydraulic conductivity anisotropy. Different upper regolith hydraulic conductivity anisotropy factors had to be tested in order to describe streamflow in both catchments.

This paper describes four decades of research carried out at the University of Michigan that has investigated the clinical alteration of the transverse dimension of the face, with specific attention given to orthodontic and orthopedic treatment of patients in the early mixed dentition. The results of a series of clinical studies beginning in the late 1970s are described that consider the etiology of dental crowding as well as treatment effects produced by expansion in the late mixed/early permanent dentition and the early mixed dentition. A treatment protocol for early treatment is described that includes the lower Schwarz appliance, a bonded acrylic splint expander and the placement of anterior brackets. A series of retrospective and prospective studies are presented, including a study investigating the spontaneous improvement of Class II malocclusion following increases in the transverse dimension. The effect of maintaining or increasing anterior arch length on subsequent mandibular second molar eruption is evaluated. The ideal timing for rapid maxillary expansion is before puberty. An individual assessment of the midpalatal suture using cone-beam computed tomography (CBCT) images can be useful when making a clinical decision between conventional and surgically assisted RME, especially for adolescent and young adult patients.

Exercise induces signaling networks to improve muscle function and confer health benefits. To identify divergent and common signaling networks during and after different exercise modalities, we performed a phosphoproteomic analysis of human skeletal muscle from a cross-over intervention of endurance, sprint, and resistance exercise. This identified 5,486 phosphosites regulated during or after at least one type of exercise modality and only 420 core phosphosites common to all exercise. One of these core phosphosites was S67 on the uncharacterized protein C18ORF25, which we validated as an AMPK substrate. Mice lacking C18ORF25 have reduced skeletal muscle fiber size, exercise capacity, and muscle contractile function, and this was associated with reduced phosphorylation of contractile and Casup2+/suphandling proteins. Expression of C18ORF25 S66/67D phospho-mimetic reversed the decreased muscle force production. This work defines the divergent and canonical exercise phosphoproteome across different modalities and identifies C18ORF25 as a regulator of exercise signaling and muscle function.

We set out to establish a novel model of temporal lobe epilepsy (TLE) in a mouse. We sought to induce TLE through the injection of kainic acid (KA) into the tail vein with subsequent development of status epilepticus (SE). Using C57BL/6 mice, we implanted hippocampal EEG recording electrodes before or after injection of KA or phosphate buffered saline (PBS). Video and EEG analysis were conducted to evaluate for SE and development of recurrent seizures, the hallmark of TLE. All mice injected with KA developed SE while those who were injected with PBS did not. Of the animals injected with KA monitored for recurrent seizures following SE, 33% developed spontaneous recurrent seizures while those injected with PBS did not. Injection of KA through the tail vein of a mouse reliably and rapidly induces SE which remits spontaneously and leads to the development of TLE in a subset of mice.

Hypertrophic cardiomyopathy (HCM) is an inherited cardiovascular disorder primarily caused by mutations in the β-myosin heavy-chain gene. The proximal subfragment 2 region (S2), 126 amino acids of myosin, binds with the C0-C2 region of cardiac myosin-binding protein-C to regulate cardiac muscle contractility in a manner dependent on PKA-mediated phosphorylation. However, it is unknown if HCM-associated mutations within S2 dysregulate actomyosin dynamics by disrupting its interaction with C0-C2, ultimately leading to HCM. Herein, we study three S2 mutations known to cause HCM: R870H, E924K, and E930Δ. First, experiments using recombinant proteins, solid-phase binding, and isothermal titrating calorimetry assays independently revealed that mutant S2 proteins displayed significantly reduced binding with C0-C2. In addition, CD revealed greater instability of the coiled-coil structure in mutant S2 proteins compared with S2Wt proteins. Second, mutant S2 exhibited 5-fold greater affinity for PKA-treated C0-C2 proteins. Third, skinned papillary muscle fibers treated with mutant S2 proteins showed no change in the rate of force redevelopment as a measure of actin–myosin cross-bridge kinetics, whereas S2Wt showed increased the rate of force redevelopment. In summary, S2 and C0-C2 interaction mediated by phosphorylation is altered by mutations in S2, which augment the speed and force of contraction observed in HCM. Modulating this interaction could be a potential strategy to treat HCM in the future.

Fast skeletal myosin-binding protein-C (fMyBP-C) is one of three MyBP-C paralogs and is predominantly expressed in fast skeletal muscle. Mutations in the gene that encodes fMyBP-C, MYBPC2, are associated with distal arthrogryposis, while loss of fMyBP-C protein is associated with diseased muscle. However, the functional and structural roles of fMyBP-C in skeletal muscle remain unclear. To address this gap, we generated a homozygous fMyBP-C knockout mouse (C2(−/−)) and characterized it both in vivo and in vitro compared to wild-type mice. Ablation of fMyBP-C was benign in terms of muscle weight, fiber type, cross-sectional area, and sarcomere ultrastructure. However, grip strength and plantar flexor muscle strength were significantly decreased in C2(−/−) mice. Peak isometric tetanic force and isotonic speed of contraction were significantly reduced in isolated extensor digitorum longus (EDL) from C2(−/−) mice. Small-angle X-ray diffraction of C2(−/−) EDL muscle showed significantly increased equatorial intensity ratio during contraction, indicating a greater shift of myosin heads toward actin, while MLL4 layer line intensity was decreased at rest, indicating less ordered myosin heads. Interfilament lattice spacing increased significantly in C2(−/−) EDL muscle. Consistent with these findings, we observed a significant reduction of steady-state isometric force during Ca(2+-)activation, decreased myofilament calcium sensitivity, and sinusoidal stiffness in skinned EDL muscle fibers from C2(−/−) mice. Finally, C2(−/−) muscles displayed disruption of inflammatory and regenerative pathways, along with increased muscle damage upon mechanical overload. Together, our data suggest that fMyBP-C is essential for maximal speed and force of contraction, sarcomere integrity, and calcium sensitivity in fast-twitch muscle.

Epilepsy Aphasia Syndromes (EAS) are a spectrum of childhood epileptic, cognitive, and language disorders of unknown etiology.CNKSR2is a strong X-linked candidate gene implicated in EAS; however, there have been no studies of genetic models to dissect how its absence may lead to EAS. Here we develop a novelCnksr2KO mouse line and show that male mice exhibit increased neural activity and have spontaneous electrographic seizures.Cnksr2KO mice also display significantly increased anxiety, impaired learning and memory, and a progressive and dramatic loss of ultrasonic vocalizations. We find that Cnksr2 is expressed in cortical, striatal, and cerebellar regions and is localized at both excitatory and inhibitory postsynapses. Proteomics analysis reveals Cnksr2 anchors key binding partners at synapses, and its loss results in significant alterations of the synaptic proteome, including proteins implicated in epilepsy disorders. Our results validate that loss of CNKSR2 leads to EAS and highlights the roles of Cnksr2 in synaptic organization and neuronal network activity.SIGNIFICANCE STATEMENTEpilepsy Aphasia Syndromes (EAS) are at the severe end of a spectrum of cognitive-behavioral symptoms seen in childhood epilepsies, and they remain an inadequately understood disorder. The prognosis of EAS is frequently poor, and patients have life-long language and cognitive disturbances. Here we describe a genetic mouse model of EAS, based on the KO of the EAS risk geneCnksr2. We show that these mice exhibit electrophysiological and behavioral phenotypes similar to those of patients, providing an important new model for future studies of EAS. We also provide insights into the molecular disturbances downstream of Cnksr2 loss by usingin vivoquantitative proteomics tools.

We compared stable isotopes of water in plant stem (xylem) water and soil collected over a complete growing season from five well‐known long‐term study sites in northern/cold regions. These spanned a decreasing temperature gradient from Bruntland Burn (Scotland), Dorset (Canadian Shield), Dry Creek (USA), Krycklan (Sweden), to Wolf Creek (northern Canada). Xylem water was isotopically depleted compared to soil waters, most notably for deuterium. The degree to which potential soil water sources could explain the isotopic composition of xylem water was assessed quantitatively using overlapping polygons to enclose respective data sets when plotted in dual isotope space. At most sites isotopes in xylem water from angiosperms showed a strong overlap with soil water; this was not the case for gymnosperms. In most cases, xylem water composition on a given sampling day could be better explained if soil water composition was considered over longer antecedent periods spanning many months. Xylem water at most sites was usually most dissimilar to soil water in drier summer months, although sites differed in the sequence of change. Open questions remain on why a significant proportion of isotopically depleted water in plant xylem cannot be explained by soil water sources, particularly for gymnosperms. It is recommended that future research focuses on the potential for fractionation to affect water uptake at the soil‐root interface, both through effects of exchange between the vapour and liquid phases of soil water and the effects of mycorrhizal interactions. Additionally, in cold regions, evaporation and diffusion of xylem water in winter may be an important process., We compared stable isotopes of water in plant stem (xylem) water and soil collected over a complete growing season from five well‐known long‐term study sites in northern/cold regions. Xylem water was isotopically depleted compared to soil waters, most notably for deuterium. At all sites except one, water sources of angiosperms could be associated with soil water, while the sources of water uptake by gymnosperms were much less easily explained., FP7 Ideas: European Research Council http://dx.doi.org/10.13039/100011199, KAW Branch‐Point project, SITES (VR), Boise State University http://dx.doi.org/10.13039/100007233, US National Science Foundation, Leverhulme Trust through the ISO‐LAND project

The study was aimed at evaluating the sagittal and transversal inclinations of upper second molars in untreated adolescents with normal occlusion.A sample of 41 subjects (16 females, 25 males) was selected from the University of Michigan Growth Study (UMGS). Digital dental casts with fully erupted second molars in occlusion were chosen (mean age 14.9 ± 1.3 years). Digital measurements were recorded with the open source software 3D Slicer ( www.slicer.org ). The digital measurements of the UMGS sample were compared with the manual measurements collected by Andrews from his sample of untreated class I subjects with normal overbite and overjet. Two mixed effect models (sagittal and transversal inclinations) were performed. The "random effect" was represented by the subjects, while the "fixed effects" were the two compared groups, the side of the arch (right and left), and the group × side interaction. Outcome variables were sagittal and transversal inclinations of the upper second molars.The UMGS group showed a significantly greater distal crown angulation (-18.9°) with respect to the Andrews sample (0.4°, P 0.0001). As for the transversal inclination, the UMGS group exhibited significantly greater lingual crown inclination (-10.6° versus -8.0°, P = 0.0118).Fully erupted maxillary second molars in a sample of adolescent subjects with normal occlusion showed significantly greater distal and lingual inclinations when compared with Andrews' values. The finding of a distal crown inclination in contrast with Andrews' observation of a mesial crown inclination suggests that revision in tip prescription for preadjusted brackets may be considered.ZIEL: Ziel der Studie war es, die sagittale und transversale Neigung der oberen zweiten Molaren bei unbehandelten Jugendlichen mit normaler Okklusion zu evaluieren.Eine Stichprobe von 41 Probanden (16 weiblich, 25 männlich) wurde aus der University of Michigan Growth Study (UMGS) ausgewählt. Es wurden digitale Zahnabdrücke mit vollständig durchgebrochenen zweiten Molaren in Okklusion ausgesucht (Durchschnittsalter 14,9 ± 1,3 Jahre). Die digitalen Messungen erfolgten mit der Open-Source-Software 3D Slicer ( www.slicer.org ). Die digitalen Messungen der UMGS-Stichprobe wurden mit den manuellen Messungen verglichen, die Andrews an seiner Stichprobe von unbehandelten Klasse-I-Probanden mit normalem Overbite und Overjet erhoben hatte. Zwei Mixed-effect-Modelle (sagittale und transversale Neigungen) wurden durchgeführt. Der „Zufallseffekt“ wurde durch die Probanden repräsentiert, die „festen Effekte“ durch die beiden verglichenen Gruppen, die Seite des Zahnbogens (rechts und links) und die Interaktion Gruppe × Seite. Ergebnisvariablen waren sagittale und transversale Inklinationen der oberen zweiten Molaren.Die UMGS-Gruppe zeigte im Vergleich zur Andrews-Gruppe eine signifikant größere distale Kronenangulation (−18,9° vs. 0,4°, p 0,0001). Hinsichtlich der transversalen Neigung wies die UMGS-Gruppe eine signifikant größere linguale Kronenneigung auf (−10,6° vs. −8,0°, p = 0,0118).Vollständig durchgebrochene obere zweite Molaren in einer Stichprobe von jugendlichen Probanden mit normaler Okklusion zeigten im Vergleich zu den Andrews-Werten signifikant größere distale und linguale Neigungen. Der Befund einer distalen Kronenneigung im Gegensatz zu Andrews’ Beobachtung einer mesialen Kronenneigung deutet darauf hin, dass eine Überarbeitung der Tip-Vorgabe bei voreingestellten Brackets in Betracht gezogen werden kann.

Introduction This study aimed to evaluate the 3-dimensional (3D) mandibular dental changes over 42 years using the registration of digital models. Methods The sample comprised digital dental models of 8 untreated subjects (4 males and 4 females) with normal occlusion measured longitudinally at ages 17 years (T1) and 60 years (T2). Using 13 landmarks placed on the mucogingival junction, we registered the T2 model on the T1 model. Three-dimensional changes in the position of the landmarks on the buccal cusp tip of the posterior teeth and incisal edge of the central incisors were measured by 2 examiners. Registration and measurements were performed using SlicerCMF (version 3.1; http://www.slicer.org ) software. Intra- and interrater agreements were evaluated using intraclass correlation coefficients and the Bland-Altman method. One-sample t tests were used for evaluating interphase 3D dental changes (P Results Adequate intra- and interrater reproducibility was found. From T1 to T2, the mandibular teeth showed significant 3D positional changes. A significant dental eruption relative to the mucogingival junction was observed for the anterior and posterior teeth. Anteroposterior movements of mandibular teeth were not significant except for the right molar that drifted mesially. Transverse movements included slight lingual tipping at canines and premolars regions. Conclusions Dental changes in untreated normal occlusion were very slight from early to mature adulthood. The eruption of the mandibular teeth was the most consistent finding. A tendency for mesial movement of molars and lingual movement of first premolars and canines was observed in the mandible during the aging process.

Machine Learning (ML) is an emerging subfield of artificial intelligence with significant resources being applied to connect computer science, statistics, and medical problems. Currently, and even more so in the future, ML algorithms applied to the orthodontic specialty will offer sophisticated and automatic models able to process and synthesize data in ways orthodontists could never do themselves, and ultimately convert data into intelligent treatment actions. This work focuses on the usefulness of two ML methodologies, LASSO networks (Ln), and Boruta selection (Ba), to simplify information from different types of pathogenic processes leading to the worsening of skeletal Class III malocclusion. Cephalometric analyses of 144 Class III untreated subjects followed longitudinally during the growth process (4–19 years) were performed. After separating subjects into two subgroups of 116 with mild (M) and 28 with very serious (VS) unfavorable growth, cephalometric features were processed using Ba and Ln algorithms for feature selection and regularization. The selection procedure revealed the unexpected predictive importance of the combination of two often overlooked craniofacial variables, SN-PP and L1-MP angles. Ln regularization highlighted additional feature interactions between M and VS growing subjects. Thus, the appropriate removal of redundant cephalometric features from the dataset contributed to the detection of subjects affected by serious unfavorable craniofacial progression and revealed the unexpected prognostic value of some skeletal feature interactions.

OBJECTIVE This investigation evaluates the evidence of case-based reasoning (CBR) in providing additional information on the prediction of future Class III craniofacial growth. SETTINGS AND SAMPLE POPULATION The craniofacial characteristics of 104 untreated Class III subjects (7-17 years of age), monitored with two lateral cephalograms obtained during the growth process, were evaluated. MATERIALS AND METHODS Data were compared with the skeletal characteristics of subjects who showed a high degree of skeletal imbalance ('prototypes') obtained from a large data set of 1263 Class III cross-sectional subjects (7-17 years of age). RESULTS The degree of similarity of longitudinal subjects with the most unbalanced prototypes allowed the identification of subjects who would develop a subsequent unfavourable skeletal growth (accuracy: 81%). The angle between the palatal plane and the sella-nasion line (PP-SN angle) and the Wits appraisal were two additional craniofacial features involved in the early prediction of the adverse progression of the Class III skeletal imbalance. CONCLUSIONS Case-based reasoning methodology, which uses a personalized inference method, may bring additional information to approximate the skeletal progression of Class III malocclusion.

Background Hypertrophic cardiomyopathy (HCM) is a genetic disease of the heart and the most common cause of sudden cardiac death in the young. HCM is considered a disease of the sarcomere owing to the large number of mutations in genes encoding sarcomeric proteins. The riddle lies in discovering how these mutations lead to disease. As a result, treatments to prevent and/or treat HCM are limited to invasive surgical myectomies or ablations. The A31P variant of cardiac myosin binding protein-C, encoded by MYBPC3, was found to be more prevalent in a cohort of Maine Coon cats with HCM. However, other mutations in MYBPC3 and MYH7 have also been associated with HCM in cats of other breeds. In this study, we expand the spectrum of genes associated with HCM in cats. Results Next Generation Whole Genome sequencing was performed using DNA isolated from peripheral blood of a Maine Coon with cardiomyopathy that tested negative for the MYBPC3 A31P variant. Through risk stratification of variants, we identified a novel, homozygous intronic variant in cardiac troponin T (TNNT2). In silico analysis of the variant suggested that it may affect normal splicing of exon 3 of TNNT2. Both parents tested heterozygous for the mutation, but were unaffected by the disease. Echocardiography analyses revealed that the proband had shown early onset congestive heart failure, which is managed with a treatment regime including ACE and aldosterone inhibitors. Conclusion In summary, we are the first to demonstrate the association between TNNT2 mutations and HCM in felines, suggesting that this gene should be included in the testing panel of genes when performing genetic testing for HCM in cats.