Development of high sustained anti-drug antibody titers and corresponding clinical decline in a late-onset Pompe disease patient after 11+ years on enzyme replacement therapy.

A late-onset Pompe disease patient developed high sustained antibody titers (HSAT) of ≥51,200 after 11+ years on alglucosidase alfa and previous tolerance. There was a corresponding worsening of motor function and rise in urinary glucose tetrasaccharide (Glc4). Following immunomodulation therapy, HSAT were eliminated with improved clinical outcomes and biomarker trends. This report highlights the importance of continued surveillance of antibody titers and biomarkers, the negative impact of HSAT, and improved outcomes with immunomodulation therapy.

Heterozygous variants in MYH10 associated with neurodevelopmental disorders and congenital anomalies with evidence for primary cilia-dependent defects in Hedgehog signaling.

PURPOSE: Nonmuscle myosin II complexes are master regulators of actin dynamics that play essential roles during embryogenesis with vertebrates possessing 3 nonmuscle myosin II heavy chain genes, MYH9, MYH10, and MYH14. As opposed to MYH9 and MYH14, no recognizable disorder has been associated with MYH10. We sought to define the clinical characteristics and molecular mechanism of a novel autosomal dominant disorder related to MYH10. METHODS: An international collaboration identified the patient cohort. CAS9-mediated knockout cell models were used to explore the mechanism of disease pathogenesis. RESULTS: We identified a cohort of 16 individuals with heterozygous MYH10 variants presenting with a broad spectrum of neurodevelopmental disorders and variable congenital anomalies that affect most organ systems and were recapitulated in animal models of altered MYH10 activity. Variants were typically de novo missense changes with clustering observed in the motor domain. MYH10 knockout cells showed defects in primary ciliogenesis and reduced ciliary length with impaired Hedgehog signaling. MYH10 variant overexpression produced a dominant-negative effect on ciliary length. CONCLUSION: These data presented a novel genetic cause of isolated and syndromic neurodevelopmental disorders related to heterozygous variants in the MYH10 gene with implications for disrupted primary cilia length control and altered Hedgehog signaling in disease pathogenesis.

Multi-target 2D tracking method for singing humpback whales using vector sensors.

Acoustic vector sensors allow estimating the direction of travel of an acoustic wave at a single point by measuring both acoustic pressure and particle motion on orthogonal axes. In a two-dimensional plane, the location of an acoustic source can thus be determined by triangulation using the estimated azimuths from at least two vector sensors. However, when tracking multiple acoustic sources simultaneously, it becomes challenging to identify and link sequences of azimuthal measurements between sensors to their respective sources. This work illustrates how two-dimensional vector sensors, deployed off the coast of western Maui, can be used to generate azimuthal tracks from individual humpback whales singing simultaneously. Incorporating acoustic transport velocity estimates into the processing generates high-quality azimuthal tracks that can be linked between sensors by cross-correlating features of their respective azigrams, a particular time-frequency representation of sound directionality. Once the correct azimuthal track associations have been made between instruments, subsequent localization and tracking in latitude and longitude of simultaneous whales can be achieved using a minimum of two vector sensors. Two-dimensional tracks and positional uncertainties of six singing whales are presented, along with swimming speed estimates derived from a high-quality track.

Measurements of open-water arctic ocean noise directionality and transport velocity.

Measurements from bottom-mounted acoustic vector sensors, deployed seasonally between 2008 and 2014 on the shallow Beaufort Sea shelf along the Alaskan North Slope, are used to estimate the ambient sound pressure power spectral density, acoustic transport velocity of energy, and dominant azimuth between 25 and 450 Hz. Even during ice-free conditions, this region has unusual acoustic features when compared against other U.S. coastal regions. Two distinct regimes exist in the diffuse ambient noise environment: one with high pressure spectral density levels but low directionality, and another with lower spectral density levels but high directionality. The transition between the two states, which is invisible in traditional spectrograms, occurs between 73 and 79 dB re 1 µPa2/Hz at 100 Hz, with the transition region occurring at lower spectral levels at higher frequencies. Across a wide bandwidth, the high-directionality ambient noise consistently arrives from geographical azimuths between 0° and 30° from true north over multiple years and locations, with a seasonal interquartile range of 40° at low frequencies and high transport velocities. The long-term stability of this directional regime, which is believed to arise from the dominance of wind-driven sources along an east-west coastline, makes it an important feature of arctic ambient sound.

Estimating acoustic cue rates in bowhead whales, Balaena mysticetus, during their fall migration through the Alaskan Beaufort Sea.

Eight years of passive acoustic data (2007-2014) from the Beaufort Sea were used to estimate the mean cue rate (calling rate) of individual bowhead whales (Balaena mysticetus) during their fall migration along the North Slope of Alaska. Calls detected on directional acoustic recorders (DASARs) were triangulated to provide estimates of locations at times of call production, which were then translated into call densities (calls/h/km2). Various assumptions were used to convert call density into animal cue rates, including the time for whales to cross the arrays of acoustic recorders, the population size, the fraction of the migration corridor missed by the localizing array system, and the fraction of the seasonal migration missed because recorders were retrieved before the end of the migration. Taking these uncertainties into account in various combinations yielded up to 351 cue rate estimates, which summarize to a median of 1.3 calls/whale/h and an interquartile range of 0.5-5.4 calls/whale/h.

Biallelic loss-of-function variants in PLD1 cause congenital right-sided cardiac valve defects and neonatal cardiomyopathy.

Congenital heart disease is the most common type of birth defect, accounting for one-third of all congenital anomalies. Using whole-exome sequencing of 2718 patients with congenital heart disease and a search in GeneMatcher, we identified 30 patients from 21 unrelated families of different ancestries with biallelic phospholipase D1 (PLD1) variants who presented predominantly with congenital cardiac valve defects. We also associated recessive PLD1 variants with isolated neonatal cardiomyopathy. Furthermore, we established that p.I668F is a founder variant among Ashkenazi Jews (allele frequency of ~2%) and describe the phenotypic spectrum of PLD1-associated congenital heart defects. PLD1 missense variants were overrepresented in regions of the protein critical for catalytic activity, and, correspondingly, we observed a strong reduction in enzymatic activity for most of the mutant proteins in an enzymatic assay. Finally, we demonstrate that PLD1 inhibition decreased endothelial-mesenchymal transition, an established pivotal early step in valvulogenesis. In conclusion, our study provides a more detailed understanding of disease mechanisms and phenotypic expression associated with PLD1 loss of function.

Automated two-dimensional localization of underwater acoustic transient impulses using vector sensor image processing (vector sensor localization).

Detecting acoustic transients by signal-to-noise ratio (SNR) becomes problematic in nonstationary ambient noise environments characteristic of coral reefs. An alternate approach presented here uses signal directionality to automatically detect and localize transient impulsive sounds collected on underwater vector sensors spaced tens of meters apart. The procedure, which does not require precise time synchronization, first constructs time-frequency representations of both the squared acoustic pressure (spectrogram) and dominant directionality of the active intensity (azigram) on each sensor. Within each azigram, sets of time-frequency cells associated with transient energy arriving from a consistent azimuthal sector are identified. Binary image processing techniques then link sets that share similar duration and bandwidth between different sensors, after which the algorithm triangulates the source location. Unlike most passive acoustic detectors, the threshold criterion for this algorithm is bandwidth instead of pressure magnitude. Data collected from shallow coral reef environments demonstrate the algorithm's ability to detect SCUBA bubble plumes and consistent spatial distributions of somniferous fish activity. Analytical estimates and direct evaluations both yield false transient localization rates from 3% to 6% in a coral reef environment. The SNR distribution of localized pulses off Hawaii has a median of 7.7 dB and interquartile range of 7.1 dB.

Roaring and repetition: How bowhead whales adjust their call density and source level (Lombard effect) in the presence of natural and seismic airgun survey noise.

Over 500 000 automated and manual acoustic localizations, measured over seven years between 2008 and 2014, were used to examine how natural wind-driven noise and anthropogenic seismic airgun survey noise influence bowhead whale call densities (calls/km2/min) and source levels during their fall migration in the Alaskan Beaufort Sea. Noise masking effects, which confound measurements of behavioral changes, were removed using a modified point transect theory. The authors found that mean call densities generally rose with increasing continuous wind-driven noise levels. The occurrence of weak airgun pulse sounds also prompted an increase in call density equivalent to a 10-15 dB change in natural noise level, but call density then dropped substantially with increasing cumulative sound exposure level (cSEL) from received airgun pulses. At low in-band noise levels the mean source level of the acoustically-active population changed to nearly perfectly compensate for noise increases, but as noise levels increased further the mean source level failed to keep pace, reducing the population's communication space. An increase of >40 dB cSEL from seismic airgun activity led to an increase in source levels of just a few decibels. These results have implications for bowhead acoustic density estimation, and evaluations of the masking impacts of anthropogenic noise.

Ocular abnormalities in a patient with congenital disorder of glycosylation type Ig.

Background: Congenital disorders of glycosylation (CDG) are a group of hereditary multisystem disorders characterized by hypoglycosylation of glycoproteins. CDG type I results in a defect in the assembly of lipid-linkedoligosaccharides or their transfer onto nascent glycoproteins. Ocular abnormalities are common in CDG, but there is no report of detailed ophthalmologic evaluation in patients with CDG type Ig in the literature.Materials and Methods: Retrospective chart review of a case of CDG type Ig with novel variant in the associated gene: ALG12.Results: In addition to typical systemic findings of CDG, our case was found to have exotropia, bilateralcataracts, and retinitis pigmentosa with extinguished electroretinography in photopic and scotopic conditions.Conclusions: We hope to extend the understanding of ALG12-related CDG type Ig with these ophthalmologic observations.

GPT2 mutations in autosomal recessive developmental disability: extending the clinical phenotype and population prevalence estimates.

The glutamate pyruvate transaminase 2 (GPT2) gene produces a nuclear-encoded mitochondrial enzyme that catalyzes the reversible transfer of an amino group from glutamate to pyruvate, generating alanine and alpha-ketoglutarate. Recessive mutations in GPT2 have been recently identified in a new syndrome involving intellectual and developmental disability (IDD), postnatal microcephaly, and spastic paraplegia. We have identified additional families with recessive GPT2 mutations and expanded the phenotype to include small stature. GPT2 loss-of-function mutations were identified in four families, nine patients total, including: a homozygous mutation in one child [c.775T>C (p.C259R)]; compound heterozygous mutations in two siblings [c.812A>C (p.N271T)/c.1432_1433delGT (p.V478Rfs*73)]; a novel homozygous, putative splicing mutation [c.1035C>T (p.G345=)]; and finally, a recurrent mutation, previously identified in a distinct family [c.1210C>T (p.R404*)]. All patients were diagnosed with IDD. A majority of patients had remarkably small stature throughout development, many < 1st percentile for height and weight. Given the potential biological function of GPT2 in cellular growth, this phenotype is strongly suggestive of a newly identified clinical susceptibility. Further, homozygous GPT2 mutations manifested in at least 2 of 176 families with IDD (approximately 1.1%) in a Pakistani cohort, thereby representing a relatively common cause of recessive IDD in this population, with recurrence of the p.R404* mutation in this population. Based on variants in the ExAC database, we estimated that approximately 1 in 248 individuals are carriers of moderately or severely deleterious variants in GPT2.

Displaying bioacoustic directional information from sonobuoys using "azigrams".

The AN/SSQ-53 Directional Frequency Analysis and Recording (DIFAR) sonobuoy is an expendable device that can derive acoustic particle velocity along two orthogonal horizontal axes, along with acoustic pressure. This information enables computation of azimuths of low-frequency acoustic sources from a single compact sensor. The standard approach for estimating azimuth from these sensors is by conventional beamforming (i.e., adding weighted time series), but the resulting "cardioid" beampattern is imprecise, computationally expensive, and vulnerable to directional noise contamination for weak signals. Demonstrated here is an alternative multiplicative processing scheme that computes the "active intensity" of an acoustic signal to obtain the dominant directionality of a noise field as a function of time and frequency. This information is conveniently displayed as an "azigram," which is analogous to a spectrogram, but uses color to indicate azimuth instead of intensity. Data from several locations demonstrate this approach, which can be computed without demultiplexing the raw signal. Azigrams have been used to help diagnose sonobuoy issues, improve detectability, and estimate bearings of low signal-to-noise ratio signals. Azigrams may also enhance the detection and potential classification of signals embedded in directional noise fields.

Effects of tones associated with drilling activities on bowhead whale calling rates.

During summer 2012 Shell performed exploratory drilling at Sivulliq, a lease holding located in the autumn migration corridor of bowhead whales (Balaena mysticetus), northwest of Camden Bay in the Beaufort Sea. The drilling operation involved a number of vessels performing various activities, such as towing the drill rig, anchor handling, and drilling. Acoustic data were collected with six arrays of directional recorders (DASARs) deployed on the seafloor over ~7 weeks in Aug-Oct. Whale calls produced within 2 km of each DASAR were identified and localized using triangulation. A "tone index" was defined to quantify the presence and amplitude of tonal sounds from industrial machinery. The presence of airgun pulses originating from distant seismic operations was also quantified. For each 10-min period at each of the 40 recorders, the number of whale calls localized was matched with the "dose" of industrial sound received, and the relationship between calling rates and industrial sound was modeled using negative binomial regression. The analysis showed that with increasing tone levels, bowhead whale calling rates initially increased, peaked, and then decreased. This dual behavioral response is similar to that described for bowhead whales and airgun pulses in earlier work. Increasing call repetition rates can be a viable strategy for combating decreased detectability of signals arising from moderate increases in background noise. Meanwhile, as noise increases, the benefits of calling may decrease because information transfer becomes increasingly error-prone, and at some point calling may no longer be worth the effort.

Decadal-scale frequency shift of migrating bowhead whale calls in the shallow Beaufort Sea.

Automated and manual acoustic localizations of bowhead whale calls in the Beaufort Sea were used to estimate the minimum frequency attained by their highly variable FM-modulated call repertoire during seven westerly fall migrations. Analyses of 13 355 manual and 100 009 automated call localizations found that between 2008 and 2014 the proportion of calls that dipped below 75 Hz increased from 27% to 41%, shifting the mean value of the minimum frequency distribution from 94 to 84 Hz. Multivariate regression analyses using both generalized linear models and generalized estimating equations found that this frequency shift persisted even when accounting for ten other factors, including calling depth, call range, call type, noise level, signal-to-noise ratio, local water depth (site), airgun activity, and call spatial density. No single call type was responsible for the observed shift, but so-called "complex" calls experienced larger percentage downward shifts. By contrast, the call source level distribution remained stable over the same period. The observed frequency shift also could not be explained by migration corridor shifts, relative changes in call detectability between different frequency bands, long-term degradation in the automated airgun detector, physiological growth in the population, or behavioral responses to increasing population density (estimated via call density).

Mutations in EFL1, an SBDS partner, are associated with infantile pancytopenia, exocrine pancreatic insufficiency and skeletal anomalies in aShwachman-Diamond like syndrome.

BACKGROUND: For the final step of the maturation of the ribosome, the nascent 40S and 60S subunits are exported from the nucleus to the cell cytoplasm. To prevent premature association of these ribosomal subunits, eukaryotic initiation factor 6 (eIF6) binds the 60S subunit within the nucleus. Its release in the cytoplasm requires the interaction of EFL1 and SDBS proteins. In Shwachman-Diamond syndrome (SDS), a defective SDBS protein prevents eIF6 eviction, inhibiting its recycle to the nucleus and subsequent formation of the active 80S ribosome. OBJECTIVE: This study aims to identify the molecular basis of an SDS-like disease, manifested by pancytopenia, exocrine pancreatic insufficiency and skeletal abnormalities in six patients from three unrelated families. METHODS: Whole exome analysis was used for mutation identification. Fluorescence microscopy studies assessed the localisation of Tif6-GFP, the yeast eIF6 homologue, in yeast WT and mutant cells. Human and yeast EFL1 proteins, WT and mutants, were expressed in Saccharomyces cerevisiae BCY123 strain, and circular dichroism and small-angle X-ray scattering were used to assess the folding and flexibility of these proteins. Green malachite colorimetric assay was performed to determine the GTPase activity of WT and Efl1 mutants. RESULTS: Four patients were homozygous for p.R1095Q variant and two patients were homozygous for p.M882K variant in EFL1. Residue R1095 and M882 are conserved across species. Neither the GTPase activity of the mutant proteins nor its activation by the SDBD protein or the 60S ribosomal subunit were affected. Complementation of efl1Δ yeast cells with the EFL1 mutants rescued the slow growth phenotype. Nonetheless, Tif6-GFP was relocalised to the cytoplasm in mutant yeast cells in contrast to its nuclear localisation in WT cells. CONCLUSIONS: Mutations in EFL1 clinically manifest as SDS-like phenotype. Similar to the molecular pathology of SDS, mutant EFL1 proteins do not promote the release of cytoplasmic Tif6 from the 60S subunit, likely preventing the formation of mature ribosomes.

Acoustic vector sensor beamforming reduces masking from underwater industrial noise during passive monitoring.

Masking from industrial noise can hamper the ability to detect marine mammal sounds near industrial operations, whenever conventional (pressure sensor) hydrophones are used for passive acoustic monitoring. Using data collected from an autonomous recorder with directional capabilities (Directional Autonomous Seafloor Acoustic Recorder), deployed 4.1 km from an arctic drilling site in 2012, the authors demonstrate how conventional beamforming on an acoustic vector sensor can be used to suppress noise arriving from a narrow sector of geographic azimuths. Improvements in signal-to-noise ratio of up to 15 dB are demonstrated on bowhead whale calls, which were otherwise undetectable using conventional hydrophones.

Source level and calling depth distributions of migrating bowhead whale calls in the shallow Beaufort Sea.

Automated and manual acoustic localizations of migrating bowhead whales were used to estimate source level and calling depth distributions of their frequency-modulated-modulated calls over seven years between 2008 and 2014. Whale positions were initially triangulated using directional autonomous seafloor acoustic recorders, deployed between 25 and 55 m water depth near Kaktovik, Alaska, during the fall westward migration. Calling depths were estimated by minimizing the "discrepancy" between source level estimates from at least three recorders detecting the same call. Applying a detailed waveguide propagation model to the data yielded broadband source levels of 161 ± 9 dB re 1 µPa2 s at 1 m (SEL) for calls received between 20 and 170 Hz. Applying a simpler 15 log10(R) power-law propagation model yielded SEL source levels of 158 ± 10 dB. The most probable calling depths lay between 22 and 30 m: optimal depths for long-range acoustic signal transmission in this particular environment.

Effects of airgun sounds on bowhead whale calling rates: evidence for two behavioral thresholds.

In proximity to seismic operations, bowhead whales (Balaena mysticetus) decrease their calling rates. Here, we investigate the transition from normal calling behavior to decreased calling and identify two threshold levels of received sound from airgun pulses at which calling behavior changes. Data were collected in August-October 2007-2010, during the westward autumn migration in the Alaskan Beaufort Sea. Up to 40 directional acoustic recorders (DASARs) were deployed at five sites offshore of the Alaskan North Slope. Using triangulation, whale calls localized within 2 km of each DASAR were identified and tallied every 10 minutes each season, so that the detected call rate could be interpreted as the actual call production rate. Moreover, airgun pulses were identified on each DASAR, analyzed, and a cumulative sound exposure level was computed for each 10-min period each season (CSEL10-min). A Poisson regression model was used to examine the relationship between the received CSEL10-min from airguns and the number of detected bowhead calls. Calling rates increased as soon as airgun pulses were detectable, compared to calling rates in the absence of airgun pulses. After the initial increase, calling rates leveled off at a received CSEL10-min of ~94 dB re 1 µPa2-s (the lower threshold). In contrast, once CSEL10-min exceeded ~127 dB re 1 µPa2-s (the upper threshold), whale calling rates began decreasing, and when CSEL10-min values were above ~160 dB re 1 µPa2-s, the whales were virtually silent.

Successful reduction of high-sustained anti-idursulfase antibody titers by immune modulation therapy in a patient with severe mucopolysaccharidosis type II.

We report on a 6 year old boy with severe MPS II undergoing immune modulation therapy due to high IgG antibody titers to IV idursulfase and no significant decline in urinary GAG levels since initiating enzyme replacement therapy. He has complete deficiency of iduronate-2-sulfatase activity due to a submicroscopic deletion of the X chromosome involving the entire I2S gene but not including in the fragile X locus. At 19 months of age, IV idursulfase therapy at the recommended dose of 0.5 mg/kg/week was initiated and then increased to 1.0 mg/kg/week after no observed clinical improvement and no decline in urine GAG level. After one year of ERT at the increased dose, he had no significant decline in urinary GAG excretion and increase of anti-idursulfase IgG antibody titers to 102,000 with complete neutralizing antibodies. In light of the evidence of lack of efficacy of idursulfase therapy, the patient was started on an immune modulation regimen consisting of ofatumumab, bortezomib, methotrexate and IVIG for a 12 week period. Only a slight decrease in IgG titers and urine GAG levels was observed, leading to increased intensity of bortezomib administration and addition of dexamethasone to the regimen, while continuing with the current schedule ofatumumab, IVIG and methotrexate. Over 18 month period of immune modulation therapy, we observed a significant reduction in anti-idursulfase IgG titers and a moderate reduction in urine GAG levels compared to baseline. Modest clinical improvements were observed. Our experience suggests that future MPS II patients with a complete gene deletion may be likely to develop persistent anti-idursulfase antibody titers and may benefit from immune modulation therapy prior to the development of high titer levels.

Mouse model implicates GNB3 duplication in a childhood obesity syndrome.

Obesity is a highly heritable condition and a risk factor for other diseases, including type 2 diabetes, cardiovascular disease, hypertension, and cancer. Recently, genomic copy number variation (CNV) has been implicated in cases of early onset obesity that may be comorbid with intellectual disability. Here, we describe a recurrent CNV that causes a syndrome associated with intellectual disability, seizures, macrocephaly, and obesity. This unbalanced chromosome translocation leads to duplication of over 100 genes on chromosome 12, including the obesity candidate gene G protein ß3 (GNB3). We generated a transgenic mouse model that carries an extra copy of GNB3, weighs significantly more than its wild-type littermates, and has excess intraabdominal fat accumulation. GNB3 is highly expressed in the brain, consistent with G-protein signaling involved in satiety and/or metabolism. These functional data connect GNB3 duplication and overexpression to elevated body mass index and provide evidence for a genetic syndrome caused by a recurrent CNV.

High dose genistein aglycone therapy is safe in patients with mucopolysaccharidoses involving the central nervous system.

Genistein (4,5,7-trihydroexyisoflavone), a soy derived isoflavone, has been proposed as a substrate reduction therapy in patients with mucopolysaccharidoses (MPS) disorders with central nervous system involvement based on studies in cultured fibroblasts demonstrating that this agent inhibits glycosaminoglycan synthesis. Several studies have reported treatment of MPS III patients with low dose genistein (5-15mg/kg/day) with no serious adverse effects and variable neurocognitive outcomes. Mice with MPS IIIB treated with high dose (160mg/kg/day) genistein exhibited a significant decrease in heparan sulfate accumulation and neuroinflammation in the brain and improvement of the behavioral phenotype. No study to date has been performed using high dose genistein treatment in MPS patients. We initiated an open label study to assess the safety of high dose genistein treatment in MPS patients with neurological impairment. Twenty-two eligible patients were treated at least 12months with pure synthetic genistein at a dose of 150mg/kg/day. Safety labs, urine GAG levels, clinical status and history of adverse events were obtained every 3months and physical examination was performed by single examiner at least every 12months. While neurocognitive level was not a primary endpoint, participants were asked to obtain annual neurocognitive testing if available and a 9 point disability scale (FPSS) was recorded at each study visit. In the course of 12months of treatment, we observed no serious adverse events that were unexplained by the underlying condition and no severe adverse events that were felt to be potentially related to the genistein therapy. Two male subjects developed Tanner II breast development not present at baseline which could be related to the mild estrogenic effects of genistein. We observed no consistent decline in urine GAG levels; however, urinary GAG excretion was erratic. After 12months, the FPSS remained unchanged in 16 patients and declined by 1 point in 2 patients. Based on the results obtained so far, high dose oral genistein therapy appears to be safe in MPS patients and additional testing in a larger randomized placebo controlled trial is needed to further assess safety and efficacy.