International meeting on Wolf-Hirschhorn syndrome: Update on the nosology and new insights on the pathogenic mechanisms for seizures and growth delay.

"An International Meeting on Wolf-Hirschhorn Syndrome (WHS)" was held at The University Hospital La Paz in Madrid, Spain (October 13-14, 2017). One hundred and twenty-five people, including physicians, scientists and affected families, attended the meeting. Parent and patient advocates from the Spanish Association of WHS opened the meeting with a panel discussion to set the stage regarding their hopes and expectations for therapeutic advances. In keeping with the theme on therapeutic development, the sessions followed a progression from description of the phenotype and definition of therapeutic endpoints, to definition of genomic changes. These proceedings will review the major points of discussion.

A survey of antiepileptic drug responses identifies drugs with potential efficacy for seizure control in Wolf-Hirschhorn syndrome.

Seizures are present in over 90% of infants and children with Wolf-Hirschhorn syndrome (WHS). When present, they significantly affect quality of life. The goal of this study was to use caregiver reports to describe the comparative efficacies of commonly used antiepileptic medications in a large population of individuals with WHS. A web-based, confidential caregiver survey was developed to capture seizure semiology and a chronologic record of seizure treatments as well as responses to each treatment. Adverse events for each drug were also cataloged. We received 141 complete survey responses (47% response rate) describing the seizures of individuals ranging in age from 4months to 61years (90 females: 51 males). Using the Early Childhood Epilepsy Severity Scale (E-Chess), WHS-associated seizures are demonstrably severe regardless of deletion size. The best-performing antiepileptic drugs (AEDs) for controlling seizures in this cohort were broad spectrum drugs clobazam, levetiracetam, and lamotrigine; whereas, the three commonly used carboxamide class drugs: carbamazepine, phenytoin, and oxcarbazepine, were reported to have little effect on, or even exacerbate, seizures. The carboxamide class drugs, along with phenobarbital and topiramate, were also associated with the highest rate of intolerance due to cooccurrence of adverse events. Levetiracetam, clobazam, and clonazepam demonstrated higher tolerability and comparatively less severe adverse events (Wilcoxon rank sum comparison between performance of levetiracetam and carboxamide class drugs gives a p<0.0001 after multiple comparison adjustment). This is the largest survey to date assessing WHS seizures. This study design is susceptible to possible bias, as the data are largely drawn from caregiver report and investigators had limited access to medical records. Despite this, our data suggest that the genetic etiology of seizures, together with an accurate electroclinical delineation, are important components of drug selection, even in contiguous gene syndromes which may have complex seizure etiologies.

A novel germline PIGA mutation in Ferro-Cerebro-Cutaneous syndrome: a neurodegenerative X-linked epileptic encephalopathy with systemic iron-overload.

Three related males presented with a newly recognized x-linked syndrome associated with neurodegeneration, cutaneous abnormalities, and systemic iron overload. Linkage studies demonstrated that they shared a haplotype on Xp21.3-Xp22.2 and exome sequencing was used to identify candidate variants. Of the segregating variants, only a PIGA mutation segregated with disease in the family. The c.328_330delCCT PIGA variant predicts, p.Leu110del (or c.1030_1032delCTT, p.Leu344del depending on the reference sequence). The unaffected great-grandfather shared his X allele with the proband but he did not have the PIGA mutation, indicating that the mutation arose de novo in his daughter. A single family with a germline PIGA mutation has been reported; affected males had a phenotype characterized by multiple congenital anomalies and severe neurologic impairment resulting in infantile lethality. In contrast, affected boys in the family described here were born without anomalies and were neurologically normal prior to onset of seizures after 6 months of age, with two surviving to the second decade. PIGA encodes an enzyme in the GPI anchor biosynthesis pathway. An affected individual in the family studied here was deficient in GPI anchor proteins on granulocytes but not erythrocytes. In conclusion, the PIGA mutation in this family likely causes a reduction in GPI anchor protein cell surface expression in various cell types, resulting in the observed pleiotropic phenotype involving central nervous system, skin, and iron metabolism.

Perceptions of equine-assisted activities and therapies by parents and children with spinal muscular atrophy.

PURPOSE: To identify the physical and psychosocial effects of equine-assisted activities and therapies (EAATs) on children with spinal muscular atrophy (SMA) from the perspective of the children and their parents. METHODS: The families of all eligible children with SMA, who reported participation in EAAT, from a Western metropolitan academic center were contacted and invited to participate. This study implemented qualitative, semistructured interviews of children with SMA and their parents. RESULTS: Three themes emerged from the qualitative content analysis: physical/psychosocial benefits; relationship development with the horses, instructors, and children; and barriers to continued EAAT engagement. CONCLUSIONS: The data suggest that the overall EAAT experience was a source of enjoyment, self-confidence, and normalcy for the children with SMA. The results of this study provide preliminary support for the use of EAAT among children with SMA.

Allied Dental Students' Perceived Knowledge, Confidence, and Attitudes Regarding Disaster Victim Identification Topics.

Purpose Disaster victim identification (DVI) service requires knowledge, confidence, and an attitude (KCA) of readiness. The purpose of this study was to assess allied dental students' perceived KCA regarding DVI skills and topics.Methods A convenience sample of senior dental hygiene students (n=27) and senior dental assistant students (n=14) were recruited by email then presented mismatched simulated antemortem (AM) and postmortem (PM) bitewing radiographs and asked to indicate correct matches. Collectively, participants made 205 radiographic matches and indicated 205 degrees of certainty binarily as "positive" or "possible" (one per match). Participants also completed a researcher designed pretest/posttest electronic survey with seven 3-point Likert-scale items with answer options of "slightly", "moderately", or "extremely" regarding self-perceived knowledge. Statistical analyses were conducted with R software using an α=0.05 significance level.Results A total of n=41 students participated, yielding a response rate of 85.4%. A one-sided linear trend test revealed statistically significant increases of perceived confidence in knowledge from pretest to posttest regarding forensic odontologists' role in DVI (p<0.0001), DVI applications for mass fatality incidents (MFI) (p<0.0001), role of dental radiology in DVI (p<0.0001), and dental morphology applications for DVI (p<0.0001). Participants indicated moderate or extreme confidence in personal clinical skillsets to assist forensic odontologists with DVI. A one-sided Fisher's exact test revealed a statistically significant (p<0.05) positive association between expressed degree of certainty (confidence) and correct radiographic matches. A one-sided linear trend test revealed statistically significant (p<0.0001) improvements in attitude regarding participants' perceived importance for their respective professions to volunteer in DVI.Conclusion Participants of this study reported significant improvements of self-perceived KCA regarding DVI skills and topics. These characteristics may encourage allied dental professionals to pursue further DVI educational opportunities and future service when support is needed for MFI.

Dental hygiene students' matching accuracy when comparing antemortem dental radiographs and oral photographs to simulated postmortem WinID3® odontograms.

Matching dental antemortem (AM) and postmortem (PM) data for human identification is especially challenging when the workforce is limited. Dental hygienists have served mass fatality incidents (MFIs) due to dental-related expertise. However, forensics within dental hygiene education and research on transferable skills is limited. This qualitative balance design study assessed senior dental hygiene students' match accuracy of simulated cases varying in dental identifiers based on AM full mouth series (FMS) radiographs and oral photographs to PM WinID3® odontograms to demonstrate possible disaster victim identification (DVI) transferable skills gained during formal education. A convenience sample of senior dental hygiene students (n = 31) was presented information on WinID3® interpretation, then presented with 5 mismatched cases and asked to visually interpret each to make 10 total matches; five based on AM FMS with simulated PM WinID3® odontograms and five based on AM photographs with PM WinID3® odontograms. Match accuracy scores ranged from 41.9% to 58.1% for cases with 1-10 identifiers, and 77.4% to 93.5% for cases with 11-40 identifiers. Accuracy when matching AM radiographs to PM odontograms versus AM photographs to PM odontograms was compared and revealed no statistical differences in match accuracy depending on image type (p = 0.388 to 1.000). Results of this pilot study suggests transferable match accuracy skills resulted from the participants' dental hygiene formal education. These baseline skills with additional specialized training support the rationale for dental hygienists serving on DVI teams. More research is needed in education and practice when preparing dental hygienists for forensic-based service.

Dental Hygiene and Dental Assistant Students' Simulated DVI Radiographic Match Accuracy: A pilot study.

Purpose Allied dental health care professionals have served on disaster victim identification (DVI) teams; however, the literature is void of statistical measures regarding transferable skills and disaster preparedness. The purpose of this study was to assess second year dental hygiene and dental assistant students' match accuracy for simulated DVI radiographs and compare the match accuracy between the student groups.Methods Five patient cases were chosen at random to retrospectively collect sets of digital bitewing radiographs from two time periods. The five retrospectively selected sets of images served as simulated antemortem (AM) and postmortem (PM) radiographs. A convenience sample of second year dental hygiene and dental assistant students from two institutions (n=48) were invited to participate in this IRB-exempt descriptive observational study. The previously selected AM and PM images were randomly mismatched, and participants were asked to visually compare the image sets and indicate the matches using a drag and drop feature in an electronic survey instrument. Descriptive statistics were used to analyze the data; the significance level was set at α=0.05.Results A total of 41 dental hygiene and dental assistant students agreed to participate for a response rate of 85.4%. Eighty-five percent of the participants accurately matched five out of five sets while the remaining 15% accurately matched three out of five sets. A one-sample binomial proportion test revealed that 80% of the participants were able to match at least four out of five sets (p<0.001). Dental hygiene students demonstrated increased matching performance as compared to dental assisting students (p=0.013).Conclusion Both dental hygiene and dental assistant students demonstrated transferable DVI skills to accurately match simulated AM and PM radiographs. Future research is needed in a larger sample to develop and assess best practices of DVI training to build on existing skills for allied dental health care professionals.

New therapeutic approaches to spinal muscular atrophy.

Bench to bedside progress has been widely anticipated for a growing number of neurodegenerative disorders. Of these, spinal muscular atrophy (SMA) is perhaps the best poised to capitalize on advances in targeted therapeutics development over the next few years. Several laboratories have achieved compelling success in SMA animal models using sophisticated methods for targeted delivery, repair, or increased expression of the survival motor neuron protein, SMN. The clinical community is actively collaborating to identify, develop, and validate outcome measures and biomarkers in parallel with laboratory efforts. Innovative trial design and synergistic approaches to maximize proactive care in conjunction with treatment with one or more of the promising pharmacologic and biologic therapies currently in the pipeline will maximize our chances to achieve meaningful outcomes for patients. This review highlights recent promising scientific and clinical advances bringing us ever closer to effective treatment(s) for our patients with SMA.

Population-Based Prevalence of Myotonic Dystrophy Type 1 Using Genetic Analysis of Statewide Blood Screening Program.

OBJECTIVE: To determine whether the genetic prevalence of the CTG expansion in the DMPK gene associated with myotonic dystrophy type 1 (DM1) in an unbiased cohort is higher than previously reported population estimates, ranging from 5 to 20 per 100,000 individuals. METHODS: This study used a cross-sectional cohort of deidentified dried blood spots from the newborn screening program in the state of New York, taken from consecutive births from 2013 to 2014. Blood spots were screened for the CTG repeat expansion in the DMPK gene using triplet-repeat primed PCR and melt curve analysis. Melt curve morphology was assessed by 4 blinded reviewers to identify samples with possible CTG expansion. Expansion of the CTG repeat was validated by PCR fragment sizing using capillary electrophoresis for samples classified as positive or premutation to confirm the result. Prevalence was calculated as the number of samples with CTG repeat size ≥50 repeats compared to the overall cohort. RESULTS: Of 50,382 consecutive births, there were 24 with a CTG repeat expansion ≥50, consistent with a diagnosis of DM1. This represents a significantly higher DM1 prevalence of 4.76 per 10,000 births (95% confidence interval 2.86-6.67) or 1 in every 2,100 births. There were an additional 96 samples (19.1 per 10,000 or 1 in 525 births) with a CTG expansion in the DMPK gene in the premutation range (CTG)35-49. CONCLUSION: The prevalence of individuals with CTG repeat expansions in DMPK is up to 5 times higher than previous reported estimates. This suggests that DM1, with multisystemic manifestations, is likely underdiagnosed in practice.

High throughput screening for expanded CTG repeats in myotonic dystrophy type 1 using melt curve analysis.

BACKGROUND: Myotonic dystrophy type 1 (DM1) is caused by CTG repeat expansions in the DMPK gene and is the most common form of muscular dystrophy. Patients can have long delays from onset to diagnosis, since clinical signs and symptoms are often nonspecific and overlapping with other disorders. Clinical genetic testing by Southern blot or triplet-primed PCR (TP-PCR) is technically challenging and cost prohibitive for population surveys. METHODS: Here, we present a high throughput, low-cost screening tool for CTG repeat expansions using TP-PCR followed by high resolution melt curve analysis with saturating concentrations of SYBR GreenER dye. RESULTS: We determined that multimodal melt profiles from the TP-PCR assay are a proxy for amplicon length stoichiometry. In a screen of 10,097 newborn blood spots, melt profile analysis accurately reflected the tri-modal distribution of common alleles from 5 to 35 CTG repeats, and identified the premutation and full expansion alleles. CONCLUSION: We demonstrate that robust detection of expanded CTG repeats in a single tube can be achieved from samples derived from specimens with minimal template DNA such as dried blood spots (DBS). This technique is readily adaptable to large-scale testing programs such as population studies and newborn screening programs.

Postmortem Dental Records Identification by Dental Hygiene Students: A pilot study.

Purpose: Dental hygienists have the potential for filling critical roles in multidisciplinary victim identification teams. The purpose of this pilot study was to evaluate the accuracy of dental hygiene students utilizing dental charting, bitewings, and skull dentitions for the purpose of making identification matches.Methods: Thirty senior dental hygiene students (n=30) independently viewed an asynchronous online multimedia-based presentation on the procedures used for collecting and recording forensic dental evidence. Following the presentation participants attempted to chart and match three bitewing radiograph sets to three human skull dentitions by correlating matches/exclusions. Immediately following the activity, each student completed a questionnaire rating the difficulty of the exercise, as well as their confidence, and willingness to volunteer as a forensics team member. Descriptive statistics were used to analyze the data.Results: Of the total sample 36.7% (n=11) reported having prior experiences with dental radiography; while the majority (63.33%, n=19) reported no prior experience. Participants' accuracy scores for dental charting ranged from 91.23% (SD=9.42) to 99.06% (SD=3.60), with no statistically significant difference based on prior experience (p>0.05). The average interrater reliability was 86% (p<0.0001), indicating a high level of agreement with charting skulls and radiographs. No statistically significant differences were found for charting time, perceived difficulty, or level of confidence when comparing experience among the participants (p>0.05).Conclusions: Regardless of previous experience, dental hygiene students were able to match postmortem dentitions and radiographs with good interrater reliability and did not differ statistically for charting time, perceived difficulty, or confidence. Results suggest dental hygienists can work as effective victim identification team members when educational programs are implemented.

Workplace Bullying: A survey of Virginia dental hygienists.

Purpose: Workplace bullying in health care has been identified as a problem that negatively affects career satisfaction, career longevity and patient outcomes. The purpose of this pilot study was to determine the prevalence of workplace bullying in a convenience sample of dental hygienists in the state of Virginia.Methods: Two hundred and forty Virginia dental hygienists attending a continuing education seminar were invited to participate. Using the Negative Acts Questionnaire-Revised (NAQ-R), respondents were asked to indicate how often they had experienced 22 negative acts or behaviors according to rate of occurrence (never, now and then or monthly, weekly or daily). Bullying was defined as experiencing two or more of the specified negative behaviors over the past 6 months. The negative behaviors were categorized into three subgroups: work-related bullying, personal bullying and physical intimidation.Results: The response rate was 64%. Data revealed almost one fourth (24%) of respondents experienced workplace bullying. The most frequent behaviors experienced by those being bullied were having their opinions and views ignored (73%), experiencing unmanageable workloads (68%) and having their work excessively monitored (68%), on a weekly or daily basis.Conclusions: Results from this study suggest approximately 1 out of 4 Virginia dental hygienists responding to this survey experience workplace bullying. Education and support to ensure identification of bullying may be helpful in promoting proactive awareness, prevention strategies and a healthier work environment leading to greater job satisfaction.

Novel PLP1 Mutations Identified With Next-Generation Sequencing Expand the Spectrum of PLP1-Associated Leukodystrophy Clinical Phenotypes.

Next-generation sequencing was performed for 2 families with an undiagnosed neurologic disease. Analysis revealed X-linked mutations in the proteolipid protein 1 (PLP1) gene, which is associated with X-linked Pelizaeus-Merzbacher disease and Spastic Paraplegia type 2. In family A, the novel PLP1 missense mutation c.617T>A (p.M206K) was hemizygous in the 2 affected male children and heterozygous in the mother. In family B, the novel de novoPLP1 frameshift mutation c.359_369del (p.G120fs) was hemizygous in the affected male child. Although PLP1 mutations have been reported to cause an increasingly wide range of phenotypes inclusive of the dystonia, spastic paraparesis, motor neuronopathy, and leukodystrophy observed in our patients, atypical features included the cerebrospinal fluid deficiency of neurotransmitter and pterin metabolites and the delayed appearance of myelin abnormalities on neuroimaging studies. Next-generation sequencing studies provided a diagnosis for these families with complex leukodystrophy disease phenotypes, which expanded the spectrum of PLP1-associated leukodystrophy clinical phenotypes.

Whole-genome analysis for effective clinical diagnosis and gene discovery in early infantile epileptic encephalopathy.

Early infantile epileptic encephalopathy (EIEE) is a devastating epilepsy syndrome with onset in the first months of life. Although mutations in more than 50 different genes are known to cause EIEE, current diagnostic yields with gene panel tests or whole-exome sequencing are below 60%. We applied whole-genome analysis (WGA) consisting of whole-genome sequencing and comprehensive variant discovery approaches to a cohort of 14 EIEE subjects for whom prior genetic tests had not yielded a diagnosis. We identified both de novo point and INDEL mutations and de novo structural rearrangements in known EIEE genes, as well as mutations in genes not previously associated with EIEE. The detection of a pathogenic or likely pathogenic mutation in all 14 subjects demonstrates the utility of WGA to reduce the time and costs of clinical diagnosis of EIEE. While exome sequencing may have detected 12 of the 14 causal mutations, 3 of the 12 patients received non-diagnostic exome panel tests prior to genome sequencing. Thus, given the continued decline of sequencing costs, our results support the use of WGA with comprehensive variant discovery as an efficient strategy for the clinical diagnosis of EIEE and other genetic conditions.

Testing a Novel 3D Printed Radiographic Imaging Device for Use in Forensic Odontology.

There are specific challenges related to forensic dental radiology and difficulties in aligning X-ray equipment to teeth of interest. Researchers used 3D printing to create a new device, the combined holding and aiming device (CHAD), to address the positioning limitations of current dental X-ray devices. Participants (N = 24) used the CHAD, soft dental wax, and a modified external aiming device (MEAD) to determine device preference, radiographer's efficiency, and technique errors. Each participant exposed six X-rays per device for a total of 432 X-rays scored. A significant difference was found at the 0.05 level between the three devices (p = 0.0015), with the MEAD having the least amount of total errors and soft dental wax taking the least amount of time. Total errors were highest when participants used soft dental wax-both the MEAD and the CHAD performed best overall. Further research in forensic dental radiology and use of holding devices is needed.

Research conference summary from the 2014 International Task Force on ATP1A3-Related Disorders.

OBJECTIVE: ATP1A3-related neurologic disorders encompass a broad range of phenotypes that extend well beyond initial phenotypic criteria associated with alternating hemiplegia of childhood (AHC) and rapid-onset dystonia parkinsonism. METHODS: In 2014, the Alternating Hemiplegia of Childhood Foundation hosted a multidisciplinary workshop intended to address fundamental challenges surrounding the diagnosis and management of individuals with ATP1A3-related disorders. RESULTS: Workshop attendees were charged with the following: (1) to achieve consensus on expanded diagnostic criteria to facilitate the identification of additional patients, intended to supplement existing syndrome-specific diagnostic paradigms; (2) to standardize definitions for the broad range of paroxysmal manifestations associated with AHC to disseminate to families; (3) to create clinical recommendations for common recurrent issues facing families and medical care providers; (4) to review data related to the death of individuals in the Alternating Hemiplegia of Childhood Foundation database to guide future efforts in identifying at-risk subjects and potential preventative measures; and (5) to identify critical gaps where we most need to focus national and international research efforts. CONCLUSIONS: This report summarizes recommendations of the workshop committee, highlighting the key phenotypic features to facilitate the diagnosis of possible ATP1A3 mutations, providing recommendations for genetic testing, and outlining initial acute management for common recurrent clinical conditions, including epilepsy.

Reassessing carrier status for dystrophinopathies.

The cloning of the DMD gene, and the identifications of mutations in it as the cause of Duchenne muscular dystrophy (DMD), makes a compelling story that is aptly told elsewhere.1 The locus-the largest in the human genome-consists of 79 exons, distributed over 2.5 million nucleotides on the X chromosome, which are assembled into a complementary DNA (cDNA) of around 14 kb encoding the predominant muscle isoform of the dystrophin protein.2 The size of the gene, and the number of exons, had historically made mutation analysis challenging. For more than a decade, the standard clinical assay was a multiplex PCR test that amplified sequences from a limited number of exons; nevertheless, because it included exons within the deletion hotspots of the gene, this method could confirm the presence of mutations in up to 98% of boys with exonic deletions.3,4.

Assessment of Mass Fatality Preparedness and Response Content in Dental Hygiene Education.

When mass fatality incidents (MFIs) occur, they can quickly overwhelm local, state, and government agencies, resources, and personnel. It is important to have a rapid and effective response with skilled, multidisciplinary victim identification teams since specific skill sets are necessary to participate in mass fatality preparedness and response. The aims of this study were to determine the extent of formal education related to mass fatality preparedness and response training in U.S. dental hygiene programs and to assess program directors' perceptions of the need for such training. A 23-item cross-sectional survey was emailed to 319 U.S. dental hygiene programs in 2015. Survey questions addressed if the program offered mass fatality preparedness and response training to its students and how much training was given, as well as collecting respondents' demographics and opinions regarding education and training. An overall response rate of 36% was obtained, with 111 program chairs completing the survey. The results showed that only a small percentage of responding programs incorporated coursework related to mass fatality and preparedness in their curricula. Of the responding programs, 84% had no formal instruction on the role of a dental hygienist in MFIs; however, 53 of 69 program directors agreed or strongly agreed that the role of dental hygienists in MFIs should be covered in dental hygiene curricula. The top three barriers to incorporating such training reported by respondents were time requirements, lack of faculty expertise, and lack of equipment. Future research is needed to establish standardized competencies for mass fatality preparedness and response in dental hygiene education.