PRUNE1: a disease-causing gene for secondary microcephaly

In their Letter to the Editor, Karakaya et al. (2017) present an interesting case report describing the clinical course involving secondary microcephaly of a 3-year-old Turkish boy found to be homozygous for a frameshift mutation in PRUNE1 identified through whole exome sequencing. The child presented with congenital hypotonia, contractures and global developmental delay with respiratory insufficiency and seizures developing in the first year of life. The authors note that the affected child’s head circumference plotted on the 75th centile at birth, and that by 38 months of age he had developed microcephaly. Neuroimaging at 14 months revealed cerebral and cerebellar atrophy consistent with other patients described with Prune syndrome (Karaca et al., 2015; Costain et al., 2017; Zollo et al., 2017). Although the child had abnormal neurology from birth, there was a period of early developmental regression. Peripheral spasticity in the lower extremities and optic atrophy were not documented until 38 months. In addition to the PRUNE1 variant, Karakaya et al. also identified a second homozygous variant in the CCDC14 gene in the Turkish child’s whole exome sequencing data that, while listed to have an allele count of 108 in the current Genome Aggregation Database (gnomAD) release, is notably absent in homozygous fashion (Lek et al., 2016). CCDC14 is known to be expressed in human brain, reported to negatively regulate centriole duplication and interact with proteins previously associated with primary microcephaly (Firat-Karalar et al., 2014). Thus, while it seems likely that the homozygous PRUNE1 variant is primarily responsible for the clinical presentation in the Turkish child, it is impossible to determine whether there may be any phenotypical contribution from this additional homozygous sequence variant. Recently, Costain et al. (2017) described a homozygous consensus splice site variant in PRUNE1 (c.521-2A4G; NM_021222.1) in a 2-year-old Oji-Cre male who presented with congenital hypotonia and talipes, whose head circumference was large at birth ( +3 standard deviations), but by 2 years and 2 months plotted on the 50th centile, with a weight and height on the 95th and 75th centiles, respectively. However, it should be noted that the child’s father is macrocephalic ( +4 standard deviations), the published clinical photographs at 2 years 5 months of age illustrate bitemporal narrowing, a sloping forehead and large ears, consistent with a developing microcephaly, and neuroimaging revealed cortical and cerebellar atrophy. He developed respiratory insufficiency shortly after birth, and infantile spasms in the first year of life (Costain et al., 2017). It remains to be determined how the phenotypical outcomes stemming from proposed loss-of-function mutations defined by Karakaya et al. and Costain et al., relate to missense mutations published by Karaca et al. and also Zollo et al., which are likely to involve at least partial gain-of-function outcomes in PRUNE1 activity. However, as more cases are investigated and published, the phenotype associated with autosomal recessive Prune neurodevelopmental disorder, and the functional outcomes of PRUNE1 mutation, are becoming clearer. It is now apparent that while some patients have a small head at birth and others a head circumference in the normal range, the key component of the microcephaly is that it is progressive, and associated with characteristic neuroimaging findings with a thin or hypoplastic corpus callosum and cortical and cerebellar atrophy developing in early childhood. Although all patients with Prune syndrome described to date are neurologically impaired from birth, there also appears to be a neurodegenerative component with progression of the disorder. In our manuscript, we described clinical overlap of Prune syndrome with the neurodegenerative condition associated with homozygous mutations in TBCD (Zollo et al., 2017). TBCD encodes one of the five tubulin-specific chaperones that are required for a/b-tubulin de novo heterodimer formation and the disorder is characterized by developmental regression, seizures, optic atrophy and secondary microcephaly, cortical atrophy with delayed myelination, cerebellar atrophy and thinned corpus callosum (Edvardson et al., 2016; Flex et al., 2016; Miyake et al., 2016; Pode-Shakked et al., 2017). The neurodegenerative phenotype documented in the Turkish child by Karakaya et al. further demonstrates the similarities with the TBCD disorder and Prune syndrome, and confirms optic atrophy to be a feature of Prune syndrome. Interestingly, it is also becoming clear that respiratory insufficiency is a common feature of Prune syndrome, having been documented by Karakaya et al. and in the Oji-Cre child, as well as the youngest affected Omani child described in our manuscript

Dominant mutations of the Notch ligand Jagged1 cause peripheral neuropathy

Notch signaling is a highly conserved intercellular pathway with tightly regulated and pleiotropic roles in normal tissue development and homeostasis. Dysregulated Notch signaling has also been implicated in human disease, including multiple forms of cancer, and represents an emerging therapeutic target. Successful development of such therapeutics requires a detailed understanding of potential on-target toxicities. Here, we identify autosomal dominant mutations of the canonical Notch ligand Jagged1 (or JAG1) as a cause of peripheral nerve disease in 2 unrelated families with the hereditary axonal neuropathy Charcot-Marie-Tooth disease type 2 (CMT2). Affected individuals in both families exhibited severe vocal fold paresis, a rare feature of peripheral nerve disease that can be life-threatening. Our studies of mutant protein posttranslational modification and localization indicated that the mutations (p.Ser577Arg, p.Ser650Pro) impair protein glycosylation and reduce JAG1 cell surface expression. Mice harboring heterozygous CMT2-associated mutations exhibited mild peripheral neuropathy, and homozygous expression resulted in embryonic lethality by midgestation. Together, our findings highlight a critical role for JAG1 in maintaining peripheral nerve integrity, particularly in the recurrent laryngeal nerve, and provide a basis for the evaluation of peripheral neuropathy as part of the clinical development of Notch pathway-modulating therapeutics.This article is freely available via Open Access. Click on the publisher URL to access it via the publisher's site.G1002279/MRC_/Medical Research Council/United Kingdom R25 NS065729/NS/NINDS NIH HHS/United States Z01 AG000949/ImNIH/Intramural NIH HHS/United States R01 NS062869/NS/NINDS NIH HHS/United States F31 NS105404/NS/NINDS NIH HHS/United Statespre-print, post-print, publisher's version/PD

Deformation of vortex patches by boundaries

The deformation of two-dimensional vortex patches in the vicinity of fluid boundaries is investigated. The presence of a boundary causes an initially circular patch of uniform vorticity to deform. Sufficiently far away from the boundary, the deformed shape is well approximated by an ellipse. This leading order elliptical deformation is investigated via the elliptic moment model of Melander, Zabusky & Styczek [M. V. Melander, N. J. Zabusky & A. S. Styczek, J. Fluid. Mech., 167, 95 (1986)]. When the boundary is straight, the centre of the elliptic patch remains at a constant distance from the boundary, and the motion is integrable. Furthermore, since the straining flow acting on the patch is constant in time, the problem is that of an elliptic vortex patch in constant strain, which was analysed by Kida [S. Kida, J. Phys. Soc. Japan, 50, 3517 (1981)]. For more complicated boundary shapes, such as a square corner, the motion is no longer integrable. Instead, there is an adiabatic invariant for the motion. This adiabatic invariant arises due to the separation in times scales between the relatively rapid time scale associated with the rotation of the patch and the slower time scale associated with the self-advection of the patch along the boundary. The interaction of a vortex patch with a circular island is also considered. Without a background flow, conservation of angular impulse implies that the motion is again integrable. The addition of an irrotational flow past the island can drive the patch towards the boundary, leading to the possibility of large deformations and breakup.Comment: 19 pages, 16 figure

Universality in solar flare and earthquake occurrence

Earthquakes and solar flares are phenomena involving huge and rapid releases of energy characterized by complex temporal occurrence. By analysing available experimental catalogs, we show that the stochastic processes underlying these apparently different phenomena have universal properties. Namely both problems exhibit the same distributions of sizes, inter-occurrence times and the same temporal clustering: we find afterflare sequences with power law temporal correlations as the Omori law for seismic sequences. The observed universality suggests a common approach to the interpretation of both phenomena in terms of the same driving physical mechanism

Perceptions of a self-management intervention for adolescents with sickle cell disease

Objective: Individuals with sickle cell disease (SCD) are at increased risk for complications from their disease during their adolescent and young adult (AYA) years. The risk of morbidity in AYAs with SCD can be decreased with improved self-management. Existing self-management interventions typically focus on one aspect of self-management (e.g., adherence) and do not address factors that activate patients (knowledge, motivation, self-efficacy, and social support) to self-manage. Sickle Cell Thrive (SCThrive) is a mixed in-person/online, technology-enhanced (use of a mobile app), group self-management intervention that targets patient activation. To determine the most clinically significant intervention components, a qualitative study was conducted. Method: Participants were 19 AYAs (Mage = 17.05) with SCD who participated in individual semistructured phone interviews after completing SCThrive. Interview content was coded using a grounded-theory approach to generate themes related to SCThrive’s feasibility, acceptability, and motivation for and impact on self-management. Results: SCThrive was reported to be highly feasible due to the mixed in-person/online format and acceptable because they learned skills to manage SCD in a group of AYAs with SCD. Action planning and pain/mood tracking appeared to be key factors in motivating AYAs for self-management. Participants reported continuing to use self-management skills post-SCThrive (self-efficacy) including applying them to other domains of their lives (e.g., educational/vocational). Conclusions: Study results provide data that can be leveraged to enhance the feasibility, acceptability, and impact of SCThrive and other self-management interventions. Findings can also inform clinical and mobile health interventions to increase self-management in this population

MNS1 variant associated with situs inversus and male infertility

Ciliopathy disorders due to abnormalities of motile cilia encompass a range of autosomal recessive conditions typified by chronic otosinopulmonary disease, infertility, situs abnormalities and hydrocephalus. Using a combination of genome-wide SNP mapping and whole exome sequencing (WES), we investigated the genetic cause of a form of situs inversus (SI) and male infertility present in multiple individuals in an extended Amish family, assuming that an autosomal recessive founder variant was responsible. This identified a single shared (2.34 Mb) region of autozygosity on chromosome 15q21.3 as the likely disease locus, in which we identified a single candidate biallelic frameshift variant in MNS1 [NM_018365.2: c.407_410del; p.(Glu136Glyfs*16)]. Genotyping of multiple family members identified randomisation of the laterality defects in other homozygous individuals, with all wild type or MNS1 c.407_410del heterozygous carriers being unaffected, consistent with an autosomal recessive mode of inheritance. This study identifies an MNS1 variant as a cause of laterality defects and male infertility in humans, mirroring findings in Mns1-deficient mice which also display male infertility and randomisation of left-right asymmetry of internal organs, confirming a crucial role for MNS1 in nodal cilia and sperm flagella formation and function.This article is freely available via Open Access. Click on the Publisher URL to access the full-text

A quantitative LC-MS/MS method for analysis of mitochondrial -specific oxysterol metabolism

Oxysterols are critical regulators of inflammation and cholesterol metabolism in cells. They are oxidation products of cholesterol and may be differentially metabolised in subcellular compartments and in biological fluids. New analytical methods are needed to improve our understanding of oxysterol trafficking and the molecular interplay between the cellular compartments required to maintain cholesterol/oxysterol homeostasis. Here we describe a method for isolation of oxysterols using solid phase extraction and quantification by liquid chromatography-mass spectrometry, applied to tissue, cells and mitochondria. We analysed five monohydroxysterols; 24(S)-hydroxycholesterol, 25-hydroxycholesterol, 27-hydroxycholesterol, 7α-hydroxycholesterol, 7 ketocholesterol and three dihydroxysterols 7α-24(S)dihydroxycholesterol, 7α-25dihydroxycholesterol, 7α-27dihydroxycholesterol by LC-MS/MS following reverse phase chromatography. Our new method, using Triton and DMSO extraction, shows improved extraction efficiency and recovery of oxysterols from cellular matrix. We validated our method by reproducibly measuring oxysterols in mouse brain tissue and showed that mice fed a high fat diet had significantly lower levels of 24S/25diOHC, 27diOHC and 7ketoOHC. We measured oxysterols in mitochondria from peripheral blood mononuclear cells and highlight the importance of rapid cell isolation to minimise effects of handling and storage conditions on oxysterol composition in clinical samples. In addition, in vitro cell culture systems, of THP-1 monocytes and neuronal-like SH-SH5Y cells, showed mitochondrial-specific oxysterol metabolism and profiles were lineage specific. In summary, we describe a robust and reproducible method validated for improved recovery, quantitative linearity and detection, reproducibility and selectivity for cellular oxysterol analysis. This method enables subcellular oxysterol metabolism to be monitored and is versatile in its application to various biological and clinical samples.This article is freely available via Open Access. Click on the Publisher URL to access it via the publisher's site.K Borah and HR Griffiths acknowledge INClusilver funded by the European Union, grant number H2020–INNOSUP‐2017‐2017 731349; NeutroCure funded by the European Union, grant number H2020-FETOPEN-01-2018-2019-2020 861878 and Faculty Research Support Fund (FRSF) fund from the University of Surrey 2019–2020. K Borah also acknowledges support of training grant 2019 Ref T022 from VALIDATE network. I Ampong, D Gao and HR Griffiths acknowledge funding from BBSRC (China Partnering Award BB/M028100/2. D Gao acknowledges funding from the National Natural Science Foundation of China(NFSC)(GrantNo.81873665).IHKD acknowledges funding from Alzheimer's research UK midlands network grant 2019. AH Crosby and EL Baple acknowledge support from the Hereditary Spastic Paraplegia Support Group and The Diamond Jubilee Doctoral Scholarship Fund.published version, accepted version, submitted versio

Of detectability and camouflage: evaluating Pollard Walk rules using a common, cryptic butterfly

Abstract Estimating distribution and abundance of species depends on the probability at which individuals are detected. Butterflies are of conservation interest worldwide, but data collected with Pollard walks—the standard for national monitoring schemes—are often analyzed assuming that changes in detectability are negligible within recommended sampling criteria. The implications of this practice remain poorly understood. Here, we evaluated the effects of sampling conditions on butterfly counts from Pollard walks using the Arctic fritillary, a common but cryptic butterfly in boreal forests of Alberta, Canada. We used an open population binomial N‐mixture model to disentangle the effects of habitat suitability and phenology on abundance of Arctic fritillaries, and its detectability by sampling different conditions of temperature, wind, cloud cover, and hour of the day. Detectability varied by one order of magnitude within the criteria recommended for Pollard walks (P varying between 0.04 and 0.45), and simulations show how sampling in suboptimal conditions increases substantially the risk of false‐absence records (e.g., false‐absences are twice as likely than true‐presences when sampling 10 Arctic fritillaries at P = 0.04). Our results suggest that the risk of false‐absences is highest for species that are poorly detectable, low in abundance, and with short flight periods. Analysis with open population binomial N‐mixture models could improve estimates of abundance and distribution for rare species of conservation interest, while providing a powerful method for assessing butterfly phenology, abundance, and behavior using counts from Pollard walks, but require more intensive sampling than conventional monitoring schemes

Severe foot lesions in dairy goats associated with digital dermatitis treponemes

Treponeme-associated foot disease has been described in cattle with digital dermatitis and sheep with contagious ovine digital dermatitis. In this study, severe foot lesions in dairy goats associated with digital dermatitis treponemes (i.e. Treponema medium, Treponema phagedenis and Treponema pedis) were characterized macroscopically, radiographically and histologically. The main macroscopic foot lesion was of extensive solar ulceration with or without exophytic papilliform hyperkeratosis. Radiographically, the distal phalanx and distal sesamoid bones were severely damaged and remodelled. Histologically, the lesion was categorized as a chronic lymphoplasmacytic, suppurative and ulcerative pododermatitis. Immunohistochemistry identified the spirochaetal microorganisms located extracellularly in the superficial horn. Study limitations mean that the treponeme bacteria could not be considered the sole or causal agents in the cases described. © 2016 The Authors

Chandra and RXTE Spectra of the Burster GS 1826-238

Using simultaneous observations from Chandra and RXTE, we investigated the LMXB GS 1826-238 with the goal of studying its spectral and timing properties. The uninterrupted Chandra observation captured 6 bursts (RXTE saw 3 of the 6), yielding a recurrence time of 3.54 +/- 0.03 hr. Using the proportional counter array on board RXTE, we made a probable detection of 611 Hz burst oscillations in the decaying phases of the bursts with an average rms signal amplitude of 4.8%. The integrated persistent emission spectrum can be described as the dual Comptonization of ~ 0.3 keV soft photons by a plasma with kT_e ~ 20 keV and an optical depth of about 2.6 (interpreted as emission from the accretion disk corona), plus the Comptonization of hotter ~ 0.8 keV seed photons by a ~ 6.8 keV plasma (interpreted as emission from or near the boundary layer). We discovered evidence for a neutral Fe K\alpha emission line, and we found interstellar Fe L_II and Fe L_III absorption features. The burst spectrum can be fit by fixing the disk Comptonization parameters to the persistent emission best-fit values, and adding a blackbody. The blackbody/seed photon temperature at the peak of the burst is ~ 1.8 keV and returns to ~ 0.8 keV over 200 s. The blackbody radius is consistent with R_bb = 10.3-11.7 km assuming a distance of 6 kpc; however, by accounting for the fraction of the surface that is obscured by the disk as a function of binary inclination, we determined the source distance must actually be near 5 kpc in order for the stellar radius to lie within the commonly assumed range of 10-12 km.Comment: Accepted for publication in ApJ; 13 pages, 6 figure